Efficient derivation of osteoprogenitor cells from induced pluripotent stem cells for bone regeneration View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2014-07-06

AUTHORS

Yoshihiro Dogaki, Sang Yang Lee, Takahiro Niikura, Takashi Iwakura, Etsuko Okumachi, Takahiro Waki, Kenichiro Kakutani, Kotaro Nishida, Ryosuke Kuroda, Masahiro Kurosaka

ABSTRACT

PurposeThere has been great interest in the use of induced pluripotent stem cells (iPSCs) in bone regenerative strategies. To generate osteoprogenitor cells from iPSCs, the most widely used protocol relies on an intermediate using embryoid body (EB) formation. We hypothesized that an osteoprogenitor cell population could be efficiently generated from iPSCs by employing a “direct-plating method” without the EB formation step.MethodsMurine iPSC colonies were dissociated with trypsin-EDTA, and obtained single cells were cultured on gelatin-coated plates in MSC medium and FGF-2. Adherent homogeneous fibroblast-like cells obtained by this direct-plating technique were termed as direct-plated cells (DPCs). Expression levels of Oct-3/4 mRNA were analysed by real-time PCR. DPCs were evaluated for cell-surface protein expression using flow cytometry. After osteogenic induction, osteogenic differentiation ability of DPCs was evaluated.ResultsThe expression level of Oct-3/4 in DPCs was significantly down-regulated compared to that observed in iPSCs, suggesting that the cells lost pluripotency. Flow cytometry analysis revealed that DPCs exhibited cell-surface antigens similar to those of bone marrow stromal cells. Furthermore, the cells proved to have a high osteogenic differentiation capacity, which was confirmed by the significant increase in alkaline phosphatase activity, the expression levels of osteogenic genes, and calcium mineralization after 14-day osteogenic induction.ConclusionsThese findings indicate that our novel direct-plating method provides a clinically applicable, simple, and labour-efficient system for generating large numbers of homogeneous iPSC-derived osteoprogenitor cells for bone regeneration. More... »

PAGES

1779-1785

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00264-014-2440-9

DOI

http://dx.doi.org/10.1007/s00264-014-2440-9

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1014362274

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24997627


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45 antigen
46 body formation
47 bone marrow stromal cells
48 bone regeneration
49 bone regenerative strategies
50 calcium mineralization
51 capacity
52 cell populations
53 cell surface antigens
54 cell surface protein expression
55 cells
56 colonies
57 cytometry
58 cytometry analysis
59 derivation
60 differentiation ability
61 differentiation capacity
62 efficient derivation
63 embryoid body formation
64 expression
65 expression levels
66 fibroblast-like cells
67 findings
68 flow cytometry
69 formation
70 formation step
71 gelatin-coated plates
72 genes
73 great interest
74 higher osteogenic differentiation capacity
75 iPSC colonies
76 increase
77 induced pluripotent stem cells
78 induction
79 interest
80 large number
81 levels
82 mRNA
83 marrow stromal cells
84 medium
85 method
86 mineralization
87 number
88 osteogenic differentiation ability
89 osteogenic differentiation capacity
90 osteogenic genes
91 osteogenic induction
92 osteoprogenitor cell population
93 osteoprogenitor cells
94 phosphatase activity
95 plate
96 pluripotency
97 pluripotent stem cells
98 population
99 protein expression
100 protocol relies
101 real-time PCR
102 regeneration
103 regenerative strategies
104 relies
105 significant increase
106 single cells
107 stem cells
108 step
109 strategies
110 stromal cells
111 system
112 technique
113 use
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