解剖学杂志2023，Vol.46Issue(5)：401-405,445,6.DOI:10.3969/j.issn.1001-1633.2023.05.007
小鼠毛乳头细胞体外分离培养方法的优化和评价
Optimization and evaluation of the method of the isolation and cultivation of mouse dermal papilla cells in vitro
摘要
Abstract
Objective:To explore an efficient and rapid method of the isolation and cultivation of mouse dermal papilla cells(DPCs)in vitro.Methods:The dermal papilla of six-week-old female C57BL/6 mice was harvested by microdissection and digested with collagenase.DPCs were cultured and passaged in medium containing 10ng/mL basic fibroblast growth factor(bFGF)in vitro.DPCs isolated with modified methods were evaluated and compared with the DPCs with traditional methods through morphological observation,cell counting,alkaline phosphatase(ALP)specific staing and ALP mRNA expression.Results:Compared with cells by the traditional method,almost all dermal papilla by the modified method adhered,migrated,and grew faster into cell confluence.The number of cells obtained was 5.85 times that of the traditional method.In DPCs by the traditional method,aggregative growth was noted after subculture at low passage numbers.After several passages,the cell morphology gradually became larger and degenerated,and less aggregative in pattern,even disappeared(5th passage).However,the morphology of the DPCs of the modified method still showed the characteristics of long spindle shape and aggregative pattern in the 1st,2nd,and 5th passage.The results also showed that the DPCs of the modified method had high ALP activity,especially in the higher passage cells(5th passage).While nearly no ALP activity was detected in DPCs by traditional method at high passage.The expression level of ALP mRNA of the DPCs of the modified method was significantly up-regulated compared with that in the traditional method,especially in the cells at the 5th passage.Conclusion:The modified method is a simple and efficient method for isolating mouse DPCs.It can provide more reliable cell source for the research of hair follicle.关键词
毛乳头细胞/显微解剖/碱性成纤维细胞生长因子/细胞培养/小鼠Key words
dermal papilla cell/microdissection/basic fibroblast growth factor/cell culture/mouse引用本文复制引用
林享玉,祝佳铭,朱亮,王哲宁,何晶..小鼠毛乳头细胞体外分离培养方法的优化和评价[J].解剖学杂志,2023,46(5):401-405,445,6.基金项目
上海市科委科技创新行动计划项目(20JC1412300) （20JC1412300）
上海市卫生健康委员会科研项目(202040027) （202040027）
国家级大学生创新活动计划项目(202110247136) （202110247136）
上海市大学生创新活动计划项目(S202014009,S202110247140) （S202014009,S202110247140）
