麝鼠香囊腺形态学观察及腺上皮体外培养初报

Morphology of Muskrat Scent Glands and Initial Culture of the Glands Epithelium

取泌香期成年麝鼠香囊腺或对非泌香期麝鼠进行人工诱导香腺发育后取材,在形态学观察的基础上,分离香囊腺上皮细胞并进行体外初步培养。结果显示,用1mg/mlⅣ型胶原酶和0.25mg/ml胰蛋白酶分次于37℃下消化组织块5h和15min,可得到大量的分散单细胞用于原代培养;用含10%胎牛血清和双抗的DME/F12培养液为基础液,在培养液中添加10ng/ml表皮生长因子(EGF),对麝鼠香囊腺上皮细胞的存活和生长具有较明显的促进作用;腺上皮细胞可进行传代培养,也可用常规方法进行冷冻保存和复苏;体外传代培养到第5代的上皮细胞仍增殖旺盛,细胞内有丰富的脂滴,细胞分泌活动旺盛。提示,通过体外分离培养麝鼠香囊腺上皮细胞,建立香囊腺干(前体)细胞系,进而用细胞工程方法大量生产人工麝鼠香是可能的。

The cystglands of matured muskrat in secreting or the cystglands induced secreting artificially were used. Based on the morphological observation, epithelial ceils of the scent glands were isolated and cultured in vitro initially. The results showed that a great quantity of dispersed ceils in single state was available for primary culture by digestion with 1mg/ml Collagenase 1V for 5 h and 0.25mg/ml Trypsin for 15min at 37%; DME/F12 containing 10% fetal bovine serum, penicillin （100U/ml）and streptomycin（100U/ ml）, as well as EGF at the concentration of 10ng,/ml, could obviously improve the survival and proliferation of these ceils; the epithelial ceils could be serial subcultivated, freezed and thawed routinely as well; after the 5th passage, the epithelial ceils were filled with many lipid dropleds, secreting and proliferating perfectly. These results suggest that it is feasible to produce artificial muskrat scent using cell engineering by isolating, culturing cystglanitd epithelial cell and furthmore establishing a musdrat cystgland epithelial cell line.