Real-time in vivo monitoring of viable stem cells implanted on biocompatible scaffolds View Full Text


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Article Info

DATE

2008-04-25

AUTHORS

Do Won Hwang, Sung June Jang, Yun Hui Kim, Hyun Joo Kim, In Kyong Shim, Jae Min Jeong, June-Key Chung, Myung Chul Lee, Seung Jin Lee, Seung U. Kim, Soonhag Kim, Dong Soo Lee

ABSTRACT

PurposeThree-dimensional fibrous scaffolds provide an environment that enhances transplanted stem cell survival in vivo and facilitates imaging their localization, viability, and growth in vivo. To assess transplanted stem cell viability on biocompatible polymer scaffolds in vivo, we developed in vivo imaging systems for evaluation of implanted viable neural stem cells (NSC) and mesenchymal stem cells (MSC) on scaffolds using luciferase or sodium/iodide symporter (NIS) genes.MethodsFirefly luciferase stably expressing-C6 cell was established (C6-Fluc). The human neural stem cell, F3, was infected with adenoviral vector carrying luciferase gene (F3-Fluc) and MSC expressing NIS controlled by ubiquitin C promoter using lentiviral vector was established by treating blasticidine for 2 weeks (MSC-NIS). Chitosan and poly l-lactic acid (PLLA) scaffolds were used for in vivo image. In vivo expression of luciferase and human NIS was examined by bioluminescence image or 99mTc-pertechnetate gamma camera image, respectively. The cell/scaffold complex was implanted into subcutaneous or abdominal area of BALB/C nude mouse. For quantitative evaluation of cell viability, regions of interest were drawn on 99mTc-pertechnetate scintigraphy by manual.ResultsThe gradual increase of luciferase activity was observed in C6-Fluc seeded with chitosan according to the increase in the number of cells. C6-Fluc/chitosan complex subcutaneously implanted into nude mice showed longitudinal bioluminescence image until 34 days. Luciferase image of abdominal-injected C6-Fluc/PLLA complex was saturated in only 14 days, showing great cell growth due to abundant nutrients. F3 cells showed well-incorporated pattern with fibrous chitosan scaffold using scanning electron microscopy. F3 infected with Ad-Fluc showed >100-fold higher luciferase activity than luciferase activity in F3. Cell-number-dependent increase of luciferase activity was shown in F3-Fluc seeded on chitosan. F3-Fluc incorporation into chitosan after abdominal injection was clearly visible on bioluminescence image up to 11 days. Radionuclide imaging showed higher uptake by MSC-NIS on PLLA scaffolds than by MSC-NIS not seeded on a scaffold. Quantitative data showed significantly better survival of MSC-NIS on PLLA scaffolds than without scaffold at 72 h post-implantation, which concurred with histologic findings.ConclusionThese results suggest that NSC-Fluc and MSC-NIS cells incorporated within polymer scaffolds can be monitored on a long-term basis by serial in vivo imaging. We believe that a biocompatible scaffold-based imaging system could be used to assess stem cell viabilities in a non-invasive way to aid the development of regenerative therapeutics. More... »

PAGES

1887

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00259-008-0751-z

DOI

http://dx.doi.org/10.1007/s00259-008-0751-z

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1004372810

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/18437378


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26 schema:description PurposeThree-dimensional fibrous scaffolds provide an environment that enhances transplanted stem cell survival in vivo and facilitates imaging their localization, viability, and growth in vivo. To assess transplanted stem cell viability on biocompatible polymer scaffolds in vivo, we developed in vivo imaging systems for evaluation of implanted viable neural stem cells (NSC) and mesenchymal stem cells (MSC) on scaffolds using luciferase or sodium/iodide symporter (NIS) genes.MethodsFirefly luciferase stably expressing-C6 cell was established (C6-Fluc). The human neural stem cell, F3, was infected with adenoviral vector carrying luciferase gene (F3-Fluc) and MSC expressing NIS controlled by ubiquitin C promoter using lentiviral vector was established by treating blasticidine for 2 weeks (MSC-NIS). Chitosan and poly l-lactic acid (PLLA) scaffolds were used for in vivo image. In vivo expression of luciferase and human NIS was examined by bioluminescence image or 99mTc-pertechnetate gamma camera image, respectively. The cell/scaffold complex was implanted into subcutaneous or abdominal area of BALB/C nude mouse. For quantitative evaluation of cell viability, regions of interest were drawn on 99mTc-pertechnetate scintigraphy by manual.ResultsThe gradual increase of luciferase activity was observed in C6-Fluc seeded with chitosan according to the increase in the number of cells. C6-Fluc/chitosan complex subcutaneously implanted into nude mice showed longitudinal bioluminescence image until 34 days. Luciferase image of abdominal-injected C6-Fluc/PLLA complex was saturated in only 14 days, showing great cell growth due to abundant nutrients. F3 cells showed well-incorporated pattern with fibrous chitosan scaffold using scanning electron microscopy. F3 infected with Ad-Fluc showed >100-fold higher luciferase activity than luciferase activity in F3. Cell-number-dependent increase of luciferase activity was shown in F3-Fluc seeded on chitosan. F3-Fluc incorporation into chitosan after abdominal injection was clearly visible on bioluminescence image up to 11 days. Radionuclide imaging showed higher uptake by MSC-NIS on PLLA scaffolds than by MSC-NIS not seeded on a scaffold. Quantitative data showed significantly better survival of MSC-NIS on PLLA scaffolds than without scaffold at 72 h post-implantation, which concurred with histologic findings.ConclusionThese results suggest that NSC-Fluc and MSC-NIS cells incorporated within polymer scaffolds can be monitored on a long-term basis by serial in vivo imaging. We believe that a biocompatible scaffold-based imaging system could be used to assess stem cell viabilities in a non-invasive way to aid the development of regenerative therapeutics.
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32 schema:keywords BALB/c nude mice
33 C promoter
34 ConclusionThese results
35 F3
36 F3 cells
37 NIS
38 PLLA scaffolds
39 abdominal area
40 abdominal injection
41 abundant nutrients
42 acid scaffolds
43 activity
44 adenoviral vector
45 area
46 basis
47 better survival
48 biocompatible polymer scaffolds
49 biocompatible scaffolds
50 bioluminescence images
51 camera images
52 cell growth
53 cell survival
54 cell viability
55 cell/scaffold complex
56 cells
57 chitosan
58 chitosan complexes
59 chitosan scaffolds
60 complexes
61 data
62 days
63 dependent increase
64 development
65 electron microscopy
66 environment
67 evaluation
68 expression
69 facilitates
70 fibrous scaffolds
71 findings
72 gamma camera images
73 genes
74 gradual increase
75 greater cell growth
76 growth
77 high uptake
78 higher luciferase activity
79 histologic findings
80 human NIS
81 human neural stem cells
82 images
83 imaging
84 imaging system
85 incorporation
86 increase
87 injection
88 interest
89 lactic acid scaffolds
90 lentiviral vectors
91 localization
92 long-term basis
93 luciferase
94 luciferase activity
95 luciferase gene
96 manual
97 mesenchymal stem cells
98 mice
99 microscopy
100 monitoring
101 neural stem cells
102 non-invasive way
103 nude mice
104 number
105 number of cells
106 nutrients
107 patterns
108 polymer scaffolds
109 promoter
110 quantitative data
111 quantitative evaluation
112 radionuclide imaging
113 regenerative therapeutics
114 region
115 region of interest
116 results
117 scaffold complex
118 scaffolds
119 scintigraphy
120 sodium/iodide symporter gene
121 stem cell survival
122 stem cell viability
123 stem cells
124 survival
125 symporter gene
126 system
127 therapeutics
128 ubiquitin C promoter
129 uptake
130 vector
131 viability
132 viable neural stem cells
133 viable stem cells
134 vivo
135 vivo expression
136 vivo images
137 vivo monitoring
138 way
139 weeks
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