中文摘要：

以0.02%胰酶4℃过夜消化,分离表皮,37℃消化30min,打成单细胞悬液,经100μg/mLⅣ型胶原处理的培养皿黏附10min,除掉未黏附的细胞,加入培养基（80%DMEM-F12＋20%FBS＋氢化可的松（25μg/mL）＋青霉素（100IU/mL）＋链霉素（100μg/mL）＋胰岛素（15μg/mL）＋转铁蛋白＋EGF（20μg/mL））培养24h,而后将此细胞消化接种到经20μg/mL丝裂霉素C处理4h的成年绒山羊成纤维细胞滋养层上,培养2周后有各种形态的克隆状细胞集落出现,用碱性磷酸酶（AKP）染色呈深黑紫色,初步判断细胞呈阳性。本研究旨在分离绒山羊皮肤干细胞,为研究干细胞分化机制,探索绒毛发育机理,培育高产高质绒毛性状奠定分子育种理论基础。

英文摘要：

The skin sample of cashmere goat was incubated in 0.02 % trypsin at 4 ℃ overnight and the epidermis was sepa- rated,then they were placed in asterile tube containing 0.25 % trypsin at 37 ℃ for 30 minutes and the dissociated cells were ad- hered with collagen type Ⅳ to culture dishes for 10 minutes. 10 minutes later, the unattached cells were removed and the rap- idly adherent epidermal cells were cultured in stem cell growth medium（80% DMEM-F12＋20% FBS＋25 μg/mL hydrocorti- sone＋100 IU/mL penicillin＋100 μg/mL streptomycin＋15 μg/mL insulin＋transferring＋20 μg/mL EGF） for 24 h. And then dissociated cells were cultured on layer cells which the adult cashmere goat fibroblast were treated with 20 μg/mL mito- mycin C for 4 h. After 2 weeks, these cells could form different shapes of clones. Dyed with AKP, it showed deep purple and cells were judged as positive.