两种培养基对羊驼毛囊干细胞生长和增殖的影响

Effect of two kinds of culture medium on the alpaca hair follicle stem cell growth and proliferation

目的比较K-SFM和DMEM/F12两种培养基对羊驼毛囊干细胞生长和增殖的影响。方法取2-3岁羊驼背部皮肤,采用联合酶消化法分离毛囊干细胞,用无血清角质化培养基(K-SFM)和自配无血清培养基(DMEM/F12)培养羊驼毛囊干细胞,比较羊驼毛囊干细胞的生长状况,用CK19、integrin-β1特异性标志物免疫细胞化学鉴定细胞。结果免疫细胞化学证实所培养的细胞为羊驼毛囊干细胞。两种培养基均能培养一定量的羊驼毛囊干细胞,培养基K-SFM有利于羊驼毛囊干细胞的增殖,细胞生长数量多,细胞克隆形成能力强,增殖速度快;DMEM/F12培养基细胞数量少,细胞活力不强,传代次数少,增殖速度慢。结论成功分离并鉴定了羊驼毛囊干细胞,建立了适于羊驼毛囊干细胞体外扩增的培养基,K-SFM培养基较自配培养基DMEM/F12更适合羊驼毛囊干细胞的体外培养。

Objective To compare the effect of K-SFM and DMEM / F12 on alpaca hair follicle stem cell growth and proliferation.Methods Skins were collected from the back of 2- 3-year-old alpaca. The alpaca hair stem cells were disassociated with collagenase and trypsin. The keratinocyte serum free medium（ K-SFM） and the self-made serum-free medium（ DMEM / F12） were used to culture the alpaca follicle stem cells,and then the growth conditions were compared. The expression of CK19 and integrin-β1 was detected in stem cells. Results The cultured cells expressed CK19 and integrin-β1 and desmin in cytoplasm,demonstrating that they were indeed alpaca hair follicle stem cells. Some alpaca hair follicle stem cells were cultured in both two kinds of medium. K-SFM was beneficial to the growth of the alpaca follicle stem cells,with a large number of the cells,the powerful ability of cellclone formation,and the rapid reproduction. After cultured with DMEM / F12,the number of the alpaca follicle stem cells was less,its vitality was lower,its passage was decreased,and its reproduction was slower. Conclusion The present study successfully isolate and identify the alpaca hair follicle stem cells,and K-SFM medium is more suitable for the culture of alpaca hair follicle stem cells in vitro.