Establishment of immortalized mesenchymal stromal cells with red fluorescence protein expression for in vivo transplantation and tracing in the rat model with traumatic brain injury.
OBJECTIVE
Human mesenchymal stromal cells (hMSC) play a crucial role in tissue engineering and regenerative medicine, and have important clinical potential for cell therapy. However, many hMSC studies have been restricted by limited cell numbers and difficult detection in vivo. To expand the lifespan, hMSC are usually immortalized by virus-mediated gene transfer. However, these genetically modified cells easily lose critical phenotypes and stable genotypes because of insertional mutagenesis.


METHODS
We used a non-viral transfection method to establish human telomerase reverse transcriptase-immortalized cord blood hMSC (hTERT-cbMSC). We also established red fluorescent protein (RFP)-expressing hTERT-cbMSC (hTERT/RFP-cbMSC) by the same non-viral transfection method, and these cells were injected into a rat model with traumatic brain injury for in vivo detection analysis.


RESULTS
The hTERT-cbMSC could grow more than 200 population doublings with a stable doubling time and maintained differentiation capacities. hTERT/RFP-cbMSC could proliferate efficiently within 2 weeks at the injury location and could be detected easily under a fluorescent microscope. Importantly, both hTERT-cbMSC and hTERT/RFP-cbMSC showed no chromosomal abnormalities by karyotype analysis and no tumor formation in severe combined immunodeficient (SCID) mice by transplantation assay.


CONCLUSIONS
We have developed immortalized cbMSC with hTERT expression and RFP expression, which will be useful tools for stem cell research and translational study.
Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan.
Chi-Jen
CJ
Hung
Chao-Ling
CL
Yao
Fu-Chou
FC
Cheng
Mei-Ling
ML
Wu
Tzu-Hao
TH
Wang
Shiaw-Min
SM
Hwang
D050156:Adipogenesis / Q000235:genetics; D000818:Animals; D001930:Brain Injuries / Q000235:genetics / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D002461:Cell Line, Transformed; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D046228:Microarray Analysis; D023421:Models, Animal; D010012:Osteogenesis / Q000235:genetics; D051381:Rats; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D019098:Telomerase; D000097573:Red Fluorescent Protein