Establishment and Cryopreservation of a Giant Panda Skin Fibroblast Cell Line
The paper describes the process of establishment and cryopreservation of a Giant Panda (Ailuropoda melanoleuca) skin fibroblast cell line.Firstly,the skin tissue was digested with 200 IU/ml collagenase type Ⅰ under 4℃.Secondly,we added 100 IU/ml penicillin and streptomycin,5 μg/ml M-plasmocin(TM) ,5 μg/ml Amphtericin B,2 mmol/L L-Glu, 10 μg/ml insulin 40 ng/ml epidermal growth factor (EGF) and 20% (the first passage) or 10% (after the first passage) fetal bovine serum (FBS) into DME/Ham's F12 and M199,and then cultured dispersive cells in DME/Ham's F12 and tissue pieces in M199.Finally,Giant Panda skin fibroblast cells were cryopreserved in phosphate balance solution (PBS) containing 0.4% bovine serum albumin (BSA),0.1 mol/L sucrose and 10% dimethyl sulphoxide ( DMSO) . Living cell percentage was 93.258% after thawing of cells half a year after cropreservation. Thawed fibroblasts still grow well in culture, showing the successful establishment and cryopreservation of a Giant Panda fibroblast cell line.
Giant Panda (Ailuropoda melanoleuca)FibroblastsCultureSkin
NETL
NSTL
万方数据
维普
