A simple and reliable protocol for long-term culture of murine bone marrow stromal (mesenchymal) stem cells that retained their in ... View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2019-12

AUTHORS

Basem M. Abdallah, Abdullah M. Alzahrani, Ashraf M. Abdel-Moneim, Nicholas Ditzel, Moustapha Kassem

ABSTRACT

Background: Bone marrow derived stromal stem cells (BMSCs) are a clonogenic cell population that is characterized by self-renewal capacity and differentiation potential into osteoblasts, and other mesenchymal cell types. Mouse BMSCs (mBMSCs) are difficult to be cultured and propagated in vitro due to their replicative senescent phenotype, heterogeneity and high contamination with plastic adherent hematopoietic progenitors (HPCs). In this study, we described long-term culture of homogenous population of mBMSCs using simple and highly reproducible approach based on frequent subculturing (FS) at fixed split ratio in the presence of basic fibroblast growth factor (bFGF). Results: Cultured mBMSCs using this protocol (mBMSCs-FS) showed long-term survival in culture > 70 population doubling (PD) and retained their characteristic surface markers and differentiation capacity into osteoblast and adipocyte lineages. When compared to the clonal bone marrow-derived cell line ST2, mBMSCs-FS displayed more enhanced osteoblast differentiation potential and responsiveness to osteogenic factors including BMPs, IGF-1, PDGF, TGFβ1,3, FGF, cAMP, Wnt3a and VEGF. In addition, unlike ST2 cells, mBMSCs-FS maintained capacity to form ectopic bone and bone marrow stroma upon in vivo transplantation in immune-compromising mice, even at high PD levels. Interestingly, by applying the same FS + bFGF protocol, we succeeded to obtain long-term cultures of primary neonatal calvarial osteoprogenitor cells (OBs) that were cultured for more than 70 PD and maintained in vitro and in vivo osteoblast differentiation capacities. Conclusions: Our data provide a simple and reliable protocol for generating long-term cultures of mBMSCs and OBs with retained high in vitro and in vivo osteoblast differentiation capacities for use in pre-clinical and molecular mechanism studies. More... »

PAGES

3

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s12575-019-0091-3

DOI

http://dx.doi.org/10.1186/s12575-019-0091-3

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1111831396

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30733647


Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
Incoming Citations Browse incoming citations for this publication using opencitations.net

JSON-LD is the canonical representation for SciGraph data.

TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

[
  {
    "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
    "about": [
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/0604", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Genetics", 
        "type": "DefinedTerm"
      }, 
      {
        "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/06", 
        "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
        "name": "Biological Sciences", 
        "type": "DefinedTerm"
      }
    ], 
    "author": [
      {
        "affiliation": {
          "alternateName": "University of Southern Denmark", 
          "id": "https://www.grid.ac/institutes/grid.10825.3e", 
          "name": [
            "Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia", 
            "Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Abdallah", 
        "givenName": "Basem M.", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "King Faisal University", 
          "id": "https://www.grid.ac/institutes/grid.412140.2", 
          "name": [
            "Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Alzahrani", 
        "givenName": "Abdullah M.", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "Alexandria University", 
          "id": "https://www.grid.ac/institutes/grid.7155.6", 
          "name": [
            "Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia", 
            "Department of Zoology, Faculty of Science, Alexandria University, Alexandria, Egypt"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Abdel-Moneim", 
        "givenName": "Ashraf M.", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "University of Southern Denmark", 
          "id": "https://www.grid.ac/institutes/grid.10825.3e", 
          "name": [
            "Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Ditzel", 
        "givenName": "Nicholas", 
        "type": "Person"
      }, 
      {
        "affiliation": {
          "alternateName": "King Saud University", 
          "id": "https://www.grid.ac/institutes/grid.56302.32", 
          "name": [
            "Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark", 
            "Department of Cellular and Molecular Medicine, DanStem (Danish Stem Cell Center), Panum Institute, University of Copenhagen, Copenhagen, Denmark", 
            "Stem Cell Unit, Department of Anatomy, Faculty of Medicine, King Saud University, Riyadh, Saudi Arabia"
          ], 
          "type": "Organization"
        }, 
        "familyName": "Kassem", 
        "givenName": "Moustapha", 
        "type": "Person"
      }
    ], 
    "citation": [
      {
        "id": "https://doi.org/10.1634/stemcells.2005-0403", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1000890142"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1634/stemcells.2005-0403", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1000890142"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-60327-169-1_12", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1003387782", 
          "https://doi.org/10.1007/978-1-60327-169-1_12"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-60327-169-1_12", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1003387782", 
          "https://doi.org/10.1007/978-1-60327-169-1_12"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.bone.2014.07.028", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1003419950"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nprot.2012.125", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1003484660", 
          "https://doi.org/10.1038/nprot.2012.125"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.bone.2008.03.011", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1010459783"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/nprot.2008.221", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1013232651", 
          "https://doi.org/10.1038/nprot.2008.221"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1182/blood-2010-12-321539", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1014238561"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1046/j.1365-2141.2003.04669.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1015126968"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1182/blood-2007-07-103697", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1016843641"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/(sici)1097-4644(19990315)72:4<570::aid-jcb12>3.0.co;2-w", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1017222061"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.yexcr.2003.12.030", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1021532583"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.yexcr.2009.08.004", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1021556557"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1186/s13287-015-0188-9", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1024157904", 
          "https://doi.org/10.1186/s13287-015-0188-9"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.tim.2003.10.005", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1024830868"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1158/0008-5472.can-07-2665", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1027933022"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1359/jbmr.091018", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1028785954"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/s0006-291x(05)80208-6", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1029635506"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1155/2010/105940", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030139687"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1042/ba20090356", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030783904"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1182/blood-2003-09-3070", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1030824622"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-59745-104-8_6", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1033340779", 
          "https://doi.org/10.1007/978-1-59745-104-8_6"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-59745-104-8_6", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1033340779", 
          "https://doi.org/10.1007/978-1-59745-104-8_6"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1242/dev.102210", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1034533910"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1634/stemcells.21-5-527", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1035796026"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/jcb.10594", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1036014347"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/s00441-012-1458-9", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1036880772", 
          "https://doi.org/10.1007/s00441-012-1458-9"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1006/bbrc.1996.0411", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1039740315"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1006/bbrc.2000.2076", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1040512240"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.scr.2014.02.007", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1041124537"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/s0142-9612(01)00216-2", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1041423600"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/btpr.354", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1042385734"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/btpr.354", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1042385734"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1007/978-1-4939-3584-0_11", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1042628909", 
          "https://doi.org/10.1007/978-1-4939-3584-0_11"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1111/j.1440-169x.2006.00874.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1042842056"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1634/stemcells.2005-0620", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1046777701"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1161/01.cir.0000105723.91637.1c", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1046852575"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1016/j.scr.2015.09.005", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1053708092"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.18632/aging.100369", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1068670686"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "https://doi.org/10.1002/j.1460-2075.1988.tb02949.x", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1079647376"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1186/s12929-017-0321-4", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1083720013", 
          "https://doi.org/10.1186/s12929-017-0321-4"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1186/s12929-017-0321-4", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1083720013", 
          "https://doi.org/10.1186/s12929-017-0321-4"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/s41598-017-05099-1", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1090361462", 
          "https://doi.org/10.1038/s41598-017-05099-1"
        ], 
        "type": "CreativeWork"
      }, 
      {
        "id": "sg:pub.10.1038/s41598-017-13477-y", 
        "sameAs": [
          "https://app.dimensions.ai/details/publication/pub.1092170362", 
          "https://doi.org/10.1038/s41598-017-13477-y"
        ], 
        "type": "CreativeWork"
      }
    ], 
    "datePublished": "2019-12", 
    "datePublishedReg": "2019-12-01", 
    "description": "Background: Bone marrow derived stromal stem cells (BMSCs) are a clonogenic cell population that is characterized by self-renewal capacity and differentiation potential into osteoblasts, and other mesenchymal cell types. Mouse BMSCs (mBMSCs) are difficult to be cultured and propagated in vitro due to their replicative senescent phenotype, heterogeneity and high contamination with plastic adherent hematopoietic progenitors (HPCs). In this study, we described long-term culture of homogenous population of mBMSCs using simple and highly reproducible approach based on frequent subculturing (FS) at fixed split ratio in the presence of basic fibroblast growth factor (bFGF).\nResults: Cultured mBMSCs using this protocol (mBMSCs-FS) showed long-term survival in culture >\u200970 population doubling (PD) and retained their characteristic surface markers and differentiation capacity into osteoblast and adipocyte lineages. When compared to the clonal bone marrow-derived cell line ST2, mBMSCs-FS displayed more enhanced osteoblast differentiation potential and responsiveness to osteogenic factors including BMPs, IGF-1, PDGF, TGF\u03b21,3, FGF, cAMP, Wnt3a and VEGF. In addition, unlike ST2 cells, mBMSCs-FS maintained capacity to form ectopic bone and bone marrow stroma upon in vivo transplantation in immune-compromising mice, even at high PD levels. Interestingly, by applying the same FS\u2009+\u2009bFGF protocol, we succeeded to obtain long-term cultures of primary neonatal calvarial osteoprogenitor cells (OBs) that were cultured for more than 70 PD and maintained in vitro and in vivo osteoblast differentiation capacities.\nConclusions: Our data provide a simple and reliable protocol for generating long-term cultures of mBMSCs and OBs with retained high in vitro and in vivo osteoblast differentiation capacities for use in pre-clinical and molecular mechanism studies.", 
    "genre": "research_article", 
    "id": "sg:pub.10.1186/s12575-019-0091-3", 
    "inLanguage": [
      "en"
    ], 
    "isAccessibleForFree": true, 
    "isPartOf": [
      {
        "id": "sg:journal.1023701", 
        "issn": [
          "1480-9222"
        ], 
        "name": "Biological Procedures Online", 
        "type": "Periodical"
      }, 
      {
        "issueNumber": "1", 
        "type": "PublicationIssue"
      }, 
      {
        "type": "PublicationVolume", 
        "volumeNumber": "21"
      }
    ], 
    "name": "A simple and reliable protocol for long-term culture of murine bone marrow stromal (mesenchymal) stem cells that retained their in vitro and in vivo stemness in long-term culture", 
    "pagination": "3", 
    "productId": [
      {
        "name": "readcube_id", 
        "type": "PropertyValue", 
        "value": [
          "e284fcd7ac295b2e20feed56232844a72ed66dfcdc59e348b01c006b78a98e92"
        ]
      }, 
      {
        "name": "pubmed_id", 
        "type": "PropertyValue", 
        "value": [
          "30733647"
        ]
      }, 
      {
        "name": "nlm_unique_id", 
        "type": "PropertyValue", 
        "value": [
          "100963717"
        ]
      }, 
      {
        "name": "doi", 
        "type": "PropertyValue", 
        "value": [
          "10.1186/s12575-019-0091-3"
        ]
      }, 
      {
        "name": "dimensions_id", 
        "type": "PropertyValue", 
        "value": [
          "pub.1111831396"
        ]
      }
    ], 
    "sameAs": [
      "https://doi.org/10.1186/s12575-019-0091-3", 
      "https://app.dimensions.ai/details/publication/pub.1111831396"
    ], 
    "sdDataset": "articles", 
    "sdDatePublished": "2019-04-11T09:03", 
    "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
    "sdPublisher": {
      "name": "Springer Nature - SN SciGraph project", 
      "type": "Organization"
    }, 
    "sdSource": "s3://com-uberresearch-data-dimensions-target-20181106-alternative/cleanup/v134/2549eaecd7973599484d7c17b260dba0a4ecb94b/merge/v9/a6c9fde33151104705d4d7ff012ea9563521a3ce/jats-lookup/v90/0000000332_0000000332/records_121924_00000000.jsonl", 
    "type": "ScholarlyArticle", 
    "url": "https://link.springer.com/10.1186%2Fs12575-019-0091-3"
  }
]
 

Download the RDF metadata as:  json-ld nt turtle xml License info

HOW TO GET THIS DATA PROGRAMMATICALLY:

JSON-LD is a popular format for linked data which is fully compatible with JSON.

curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1186/s12575-019-0091-3'

N-Triples is a line-based linked data format ideal for batch operations.

curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1186/s12575-019-0091-3'

Turtle is a human-readable linked data format.

curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1186/s12575-019-0091-3'

RDF/XML is a standard XML format for linked data.

curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1186/s12575-019-0091-3'


 

This table displays all metadata directly associated to this object as RDF triples.

234 TRIPLES      21 PREDICATES      69 URIs      21 LITERALS      9 BLANK NODES

Subject Predicate Object
1 sg:pub.10.1186/s12575-019-0091-3 schema:about anzsrc-for:06
2 anzsrc-for:0604
3 schema:author N4d90657c6b7a4e2bbb47f831aa8ff233
4 schema:citation sg:pub.10.1007/978-1-4939-3584-0_11
5 sg:pub.10.1007/978-1-59745-104-8_6
6 sg:pub.10.1007/978-1-60327-169-1_12
7 sg:pub.10.1007/s00441-012-1458-9
8 sg:pub.10.1038/nprot.2008.221
9 sg:pub.10.1038/nprot.2012.125
10 sg:pub.10.1038/s41598-017-05099-1
11 sg:pub.10.1038/s41598-017-13477-y
12 sg:pub.10.1186/s12929-017-0321-4
13 sg:pub.10.1186/s13287-015-0188-9
14 https://doi.org/10.1002/(sici)1097-4644(19990315)72:4<570::aid-jcb12>3.0.co;2-w
15 https://doi.org/10.1002/btpr.354
16 https://doi.org/10.1002/j.1460-2075.1988.tb02949.x
17 https://doi.org/10.1002/jcb.10594
18 https://doi.org/10.1006/bbrc.1996.0411
19 https://doi.org/10.1006/bbrc.2000.2076
20 https://doi.org/10.1016/j.bone.2008.03.011
21 https://doi.org/10.1016/j.bone.2014.07.028
22 https://doi.org/10.1016/j.scr.2014.02.007
23 https://doi.org/10.1016/j.scr.2015.09.005
24 https://doi.org/10.1016/j.tim.2003.10.005
25 https://doi.org/10.1016/j.yexcr.2003.12.030
26 https://doi.org/10.1016/j.yexcr.2009.08.004
27 https://doi.org/10.1016/s0006-291x(05)80208-6
28 https://doi.org/10.1016/s0142-9612(01)00216-2
29 https://doi.org/10.1042/ba20090356
30 https://doi.org/10.1046/j.1365-2141.2003.04669.x
31 https://doi.org/10.1111/j.1440-169x.2006.00874.x
32 https://doi.org/10.1155/2010/105940
33 https://doi.org/10.1158/0008-5472.can-07-2665
34 https://doi.org/10.1161/01.cir.0000105723.91637.1c
35 https://doi.org/10.1182/blood-2003-09-3070
36 https://doi.org/10.1182/blood-2007-07-103697
37 https://doi.org/10.1182/blood-2010-12-321539
38 https://doi.org/10.1242/dev.102210
39 https://doi.org/10.1359/jbmr.091018
40 https://doi.org/10.1634/stemcells.2005-0403
41 https://doi.org/10.1634/stemcells.2005-0620
42 https://doi.org/10.1634/stemcells.21-5-527
43 https://doi.org/10.18632/aging.100369
44 schema:datePublished 2019-12
45 schema:datePublishedReg 2019-12-01
46 schema:description Background: Bone marrow derived stromal stem cells (BMSCs) are a clonogenic cell population that is characterized by self-renewal capacity and differentiation potential into osteoblasts, and other mesenchymal cell types. Mouse BMSCs (mBMSCs) are difficult to be cultured and propagated in vitro due to their replicative senescent phenotype, heterogeneity and high contamination with plastic adherent hematopoietic progenitors (HPCs). In this study, we described long-term culture of homogenous population of mBMSCs using simple and highly reproducible approach based on frequent subculturing (FS) at fixed split ratio in the presence of basic fibroblast growth factor (bFGF). Results: Cultured mBMSCs using this protocol (mBMSCs-FS) showed long-term survival in culture > 70 population doubling (PD) and retained their characteristic surface markers and differentiation capacity into osteoblast and adipocyte lineages. When compared to the clonal bone marrow-derived cell line ST2, mBMSCs-FS displayed more enhanced osteoblast differentiation potential and responsiveness to osteogenic factors including BMPs, IGF-1, PDGF, TGFβ1,3, FGF, cAMP, Wnt3a and VEGF. In addition, unlike ST2 cells, mBMSCs-FS maintained capacity to form ectopic bone and bone marrow stroma upon in vivo transplantation in immune-compromising mice, even at high PD levels. Interestingly, by applying the same FS + bFGF protocol, we succeeded to obtain long-term cultures of primary neonatal calvarial osteoprogenitor cells (OBs) that were cultured for more than 70 PD and maintained in vitro and in vivo osteoblast differentiation capacities. Conclusions: Our data provide a simple and reliable protocol for generating long-term cultures of mBMSCs and OBs with retained high in vitro and in vivo osteoblast differentiation capacities for use in pre-clinical and molecular mechanism studies.
47 schema:genre research_article
48 schema:inLanguage en
49 schema:isAccessibleForFree true
50 schema:isPartOf N71ad44a2d3bb4ba2a7a32501b699cfc4
51 N951251e0b8fc43a380a919eb2285ad39
52 sg:journal.1023701
53 schema:name A simple and reliable protocol for long-term culture of murine bone marrow stromal (mesenchymal) stem cells that retained their in vitro and in vivo stemness in long-term culture
54 schema:pagination 3
55 schema:productId N23e4dbcc80a641c58463a20e68f9963d
56 N78a56be4b56b482592c6cb69a9e155ed
57 N7a88da7f2a0a40da82047059a53d62c5
58 N8dd19101baa247a2b3ac7a54086c3058
59 Nfec20ba622564dfba957453d91a00fb4
60 schema:sameAs https://app.dimensions.ai/details/publication/pub.1111831396
61 https://doi.org/10.1186/s12575-019-0091-3
62 schema:sdDatePublished 2019-04-11T09:03
63 schema:sdLicense https://scigraph.springernature.com/explorer/license/
64 schema:sdPublisher N912b426ca94f46c0b9d276acbc56755b
65 schema:url https://link.springer.com/10.1186%2Fs12575-019-0091-3
66 sgo:license sg:explorer/license/
67 sgo:sdDataset articles
68 rdf:type schema:ScholarlyArticle
69 N23e4dbcc80a641c58463a20e68f9963d schema:name pubmed_id
70 schema:value 30733647
71 rdf:type schema:PropertyValue
72 N2ebb43b0bacc40fc99101b684233de06 schema:affiliation https://www.grid.ac/institutes/grid.10825.3e
73 schema:familyName Abdallah
74 schema:givenName Basem M.
75 rdf:type schema:Person
76 N3884e9ffbbcb4328ab42e9955d117f22 schema:affiliation https://www.grid.ac/institutes/grid.7155.6
77 schema:familyName Abdel-Moneim
78 schema:givenName Ashraf M.
79 rdf:type schema:Person
80 N49d1ff774432453fb12b9eed0bc3acfa schema:affiliation https://www.grid.ac/institutes/grid.10825.3e
81 schema:familyName Ditzel
82 schema:givenName Nicholas
83 rdf:type schema:Person
84 N4d90657c6b7a4e2bbb47f831aa8ff233 rdf:first N2ebb43b0bacc40fc99101b684233de06
85 rdf:rest Nc8b8c93b7afa49b5ae988ebeb0baa8a0
86 N586c31b11b3c444cad20ffda7bcc8267 rdf:first N49d1ff774432453fb12b9eed0bc3acfa
87 rdf:rest N690a64ac0dd5419f878303f5a89170dc
88 N690a64ac0dd5419f878303f5a89170dc rdf:first N7778326340714bcd826fa68338aa1c62
89 rdf:rest rdf:nil
90 N71ad44a2d3bb4ba2a7a32501b699cfc4 schema:volumeNumber 21
91 rdf:type schema:PublicationVolume
92 N7778326340714bcd826fa68338aa1c62 schema:affiliation https://www.grid.ac/institutes/grid.56302.32
93 schema:familyName Kassem
94 schema:givenName Moustapha
95 rdf:type schema:Person
96 N78a56be4b56b482592c6cb69a9e155ed schema:name dimensions_id
97 schema:value pub.1111831396
98 rdf:type schema:PropertyValue
99 N7a88da7f2a0a40da82047059a53d62c5 schema:name doi
100 schema:value 10.1186/s12575-019-0091-3
101 rdf:type schema:PropertyValue
102 N8dd19101baa247a2b3ac7a54086c3058 schema:name readcube_id
103 schema:value e284fcd7ac295b2e20feed56232844a72ed66dfcdc59e348b01c006b78a98e92
104 rdf:type schema:PropertyValue
105 N912b426ca94f46c0b9d276acbc56755b schema:name Springer Nature - SN SciGraph project
106 rdf:type schema:Organization
107 N951251e0b8fc43a380a919eb2285ad39 schema:issueNumber 1
108 rdf:type schema:PublicationIssue
109 Nae734bcf5d8640b8aeeeb90f97d34aa3 schema:affiliation https://www.grid.ac/institutes/grid.412140.2
110 schema:familyName Alzahrani
111 schema:givenName Abdullah M.
112 rdf:type schema:Person
113 Nb6a9707f6e5f4e91821e4b7cc7c46369 rdf:first N3884e9ffbbcb4328ab42e9955d117f22
114 rdf:rest N586c31b11b3c444cad20ffda7bcc8267
115 Nc8b8c93b7afa49b5ae988ebeb0baa8a0 rdf:first Nae734bcf5d8640b8aeeeb90f97d34aa3
116 rdf:rest Nb6a9707f6e5f4e91821e4b7cc7c46369
117 Nfec20ba622564dfba957453d91a00fb4 schema:name nlm_unique_id
118 schema:value 100963717
119 rdf:type schema:PropertyValue
120 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
121 schema:name Biological Sciences
122 rdf:type schema:DefinedTerm
123 anzsrc-for:0604 schema:inDefinedTermSet anzsrc-for:
124 schema:name Genetics
125 rdf:type schema:DefinedTerm
126 sg:journal.1023701 schema:issn 1480-9222
127 schema:name Biological Procedures Online
128 rdf:type schema:Periodical
129 sg:pub.10.1007/978-1-4939-3584-0_11 schema:sameAs https://app.dimensions.ai/details/publication/pub.1042628909
130 https://doi.org/10.1007/978-1-4939-3584-0_11
131 rdf:type schema:CreativeWork
132 sg:pub.10.1007/978-1-59745-104-8_6 schema:sameAs https://app.dimensions.ai/details/publication/pub.1033340779
133 https://doi.org/10.1007/978-1-59745-104-8_6
134 rdf:type schema:CreativeWork
135 sg:pub.10.1007/978-1-60327-169-1_12 schema:sameAs https://app.dimensions.ai/details/publication/pub.1003387782
136 https://doi.org/10.1007/978-1-60327-169-1_12
137 rdf:type schema:CreativeWork
138 sg:pub.10.1007/s00441-012-1458-9 schema:sameAs https://app.dimensions.ai/details/publication/pub.1036880772
139 https://doi.org/10.1007/s00441-012-1458-9
140 rdf:type schema:CreativeWork
141 sg:pub.10.1038/nprot.2008.221 schema:sameAs https://app.dimensions.ai/details/publication/pub.1013232651
142 https://doi.org/10.1038/nprot.2008.221
143 rdf:type schema:CreativeWork
144 sg:pub.10.1038/nprot.2012.125 schema:sameAs https://app.dimensions.ai/details/publication/pub.1003484660
145 https://doi.org/10.1038/nprot.2012.125
146 rdf:type schema:CreativeWork
147 sg:pub.10.1038/s41598-017-05099-1 schema:sameAs https://app.dimensions.ai/details/publication/pub.1090361462
148 https://doi.org/10.1038/s41598-017-05099-1
149 rdf:type schema:CreativeWork
150 sg:pub.10.1038/s41598-017-13477-y schema:sameAs https://app.dimensions.ai/details/publication/pub.1092170362
151 https://doi.org/10.1038/s41598-017-13477-y
152 rdf:type schema:CreativeWork
153 sg:pub.10.1186/s12929-017-0321-4 schema:sameAs https://app.dimensions.ai/details/publication/pub.1083720013
154 https://doi.org/10.1186/s12929-017-0321-4
155 rdf:type schema:CreativeWork
156 sg:pub.10.1186/s13287-015-0188-9 schema:sameAs https://app.dimensions.ai/details/publication/pub.1024157904
157 https://doi.org/10.1186/s13287-015-0188-9
158 rdf:type schema:CreativeWork
159 https://doi.org/10.1002/(sici)1097-4644(19990315)72:4<570::aid-jcb12>3.0.co;2-w schema:sameAs https://app.dimensions.ai/details/publication/pub.1017222061
160 rdf:type schema:CreativeWork
161 https://doi.org/10.1002/btpr.354 schema:sameAs https://app.dimensions.ai/details/publication/pub.1042385734
162 rdf:type schema:CreativeWork
163 https://doi.org/10.1002/j.1460-2075.1988.tb02949.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1079647376
164 rdf:type schema:CreativeWork
165 https://doi.org/10.1002/jcb.10594 schema:sameAs https://app.dimensions.ai/details/publication/pub.1036014347
166 rdf:type schema:CreativeWork
167 https://doi.org/10.1006/bbrc.1996.0411 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039740315
168 rdf:type schema:CreativeWork
169 https://doi.org/10.1006/bbrc.2000.2076 schema:sameAs https://app.dimensions.ai/details/publication/pub.1040512240
170 rdf:type schema:CreativeWork
171 https://doi.org/10.1016/j.bone.2008.03.011 schema:sameAs https://app.dimensions.ai/details/publication/pub.1010459783
172 rdf:type schema:CreativeWork
173 https://doi.org/10.1016/j.bone.2014.07.028 schema:sameAs https://app.dimensions.ai/details/publication/pub.1003419950
174 rdf:type schema:CreativeWork
175 https://doi.org/10.1016/j.scr.2014.02.007 schema:sameAs https://app.dimensions.ai/details/publication/pub.1041124537
176 rdf:type schema:CreativeWork
177 https://doi.org/10.1016/j.scr.2015.09.005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1053708092
178 rdf:type schema:CreativeWork
179 https://doi.org/10.1016/j.tim.2003.10.005 schema:sameAs https://app.dimensions.ai/details/publication/pub.1024830868
180 rdf:type schema:CreativeWork
181 https://doi.org/10.1016/j.yexcr.2003.12.030 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021532583
182 rdf:type schema:CreativeWork
183 https://doi.org/10.1016/j.yexcr.2009.08.004 schema:sameAs https://app.dimensions.ai/details/publication/pub.1021556557
184 rdf:type schema:CreativeWork
185 https://doi.org/10.1016/s0006-291x(05)80208-6 schema:sameAs https://app.dimensions.ai/details/publication/pub.1029635506
186 rdf:type schema:CreativeWork
187 https://doi.org/10.1016/s0142-9612(01)00216-2 schema:sameAs https://app.dimensions.ai/details/publication/pub.1041423600
188 rdf:type schema:CreativeWork
189 https://doi.org/10.1042/ba20090356 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030783904
190 rdf:type schema:CreativeWork
191 https://doi.org/10.1046/j.1365-2141.2003.04669.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1015126968
192 rdf:type schema:CreativeWork
193 https://doi.org/10.1111/j.1440-169x.2006.00874.x schema:sameAs https://app.dimensions.ai/details/publication/pub.1042842056
194 rdf:type schema:CreativeWork
195 https://doi.org/10.1155/2010/105940 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030139687
196 rdf:type schema:CreativeWork
197 https://doi.org/10.1158/0008-5472.can-07-2665 schema:sameAs https://app.dimensions.ai/details/publication/pub.1027933022
198 rdf:type schema:CreativeWork
199 https://doi.org/10.1161/01.cir.0000105723.91637.1c schema:sameAs https://app.dimensions.ai/details/publication/pub.1046852575
200 rdf:type schema:CreativeWork
201 https://doi.org/10.1182/blood-2003-09-3070 schema:sameAs https://app.dimensions.ai/details/publication/pub.1030824622
202 rdf:type schema:CreativeWork
203 https://doi.org/10.1182/blood-2007-07-103697 schema:sameAs https://app.dimensions.ai/details/publication/pub.1016843641
204 rdf:type schema:CreativeWork
205 https://doi.org/10.1182/blood-2010-12-321539 schema:sameAs https://app.dimensions.ai/details/publication/pub.1014238561
206 rdf:type schema:CreativeWork
207 https://doi.org/10.1242/dev.102210 schema:sameAs https://app.dimensions.ai/details/publication/pub.1034533910
208 rdf:type schema:CreativeWork
209 https://doi.org/10.1359/jbmr.091018 schema:sameAs https://app.dimensions.ai/details/publication/pub.1028785954
210 rdf:type schema:CreativeWork
211 https://doi.org/10.1634/stemcells.2005-0403 schema:sameAs https://app.dimensions.ai/details/publication/pub.1000890142
212 rdf:type schema:CreativeWork
213 https://doi.org/10.1634/stemcells.2005-0620 schema:sameAs https://app.dimensions.ai/details/publication/pub.1046777701
214 rdf:type schema:CreativeWork
215 https://doi.org/10.1634/stemcells.21-5-527 schema:sameAs https://app.dimensions.ai/details/publication/pub.1035796026
216 rdf:type schema:CreativeWork
217 https://doi.org/10.18632/aging.100369 schema:sameAs https://app.dimensions.ai/details/publication/pub.1068670686
218 rdf:type schema:CreativeWork
219 https://www.grid.ac/institutes/grid.10825.3e schema:alternateName University of Southern Denmark
220 schema:name Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia
221 Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark
222 rdf:type schema:Organization
223 https://www.grid.ac/institutes/grid.412140.2 schema:alternateName King Faisal University
224 schema:name Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia
225 rdf:type schema:Organization
226 https://www.grid.ac/institutes/grid.56302.32 schema:alternateName King Saud University
227 schema:name Department of Cellular and Molecular Medicine, DanStem (Danish Stem Cell Center), Panum Institute, University of Copenhagen, Copenhagen, Denmark
228 Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark
229 Stem Cell Unit, Department of Anatomy, Faculty of Medicine, King Saud University, Riyadh, Saudi Arabia
230 rdf:type schema:Organization
231 https://www.grid.ac/institutes/grid.7155.6 schema:alternateName Alexandria University
232 schema:name Biological Sciences Department, College of Science, King Faisal University, 31982, Hofuf, Al-Ahsa, Saudi Arabia
233 Department of Zoology, Faculty of Science, Alexandria University, Alexandria, Egypt
234 rdf:type schema:Organization
 




Preview window. Press ESC to close (or click here)


...