[Culturing and cryopreservation of pre-cartilaginous stem cells from neonate rat].
OBJECTIVE
To establish an effective way to cryopreserve pre-cartilaginous stem cells (PSCs) of neonate rat.


METHODS
PSCs [fibroblast growth factor-3 (FGFR-3) positive cells] were isolated and purified by magnetic cell sorting method. PSCs were cultured and amplified to the third generation. PSCs were preserved in liquid nitrogen. The biological properties of cryopreserved PSCs were investigated by reverse transcriptase polymerase chain reaction (RT-PCR), immunohistochemistry, and immunofluorescence.


RESULTS
Immunohistochemical and immunofluorescent analysis showed widespread expression of FGFR-3 in cryopreserved PSCs. FGFR-3 could be detected by RT-PCR in cryopreserved PSCs. Cryopreserved PSCs kept high cell viability, and phenotypic and proliferation characteristics of PSCs in vivo.


CONCLUSIONS
Cryopreservation of PSCs can supply adequate qualified cells for repairing the defects of epiphyseal growth plate by tissue engineering technique.
You
Hongbo
H
Department of Orthopaedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430030, PR China. youhongbo001@sina.com
Chen
Anmin
A
Cheng
Hao
H
D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D015925:Cryopreservation; D004838:Epiphyses / Q000166:cytology*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D051498:Receptor, Fibroblast Growth Factor, Type 3