用单酶消化法分离整根脐带源间充质干细胞

Isolation and characterization of mesenchymal stem cells from whole human umbilical cord with a single enzyme approach

目的 建立一种从整根脐带中分离培养脐带来源间充质干细胞（UC-MSCs）的方法.方法 用单酶消化整根脐带,并传代和扩增培养,绘制生长曲线,用流式细胞仪检测贴壁细胞表面抗原及细胞周期;在特定诱导体系中,诱导贴壁细胞向脂肪、成骨分化.结果 成功从整根脐带分离出间充质干细胞（MSCs）;生长曲线显示细胞倍增时间为（24.15±0.49）h;细胞周期G1期细胞占82.66%;贴壁细胞均表达CD93、CD105、CD44、CD29和CD73,不表达CD34、CD45、CD31和人类白细胞DR抗原（HLA-DR）;UC-MSCs能够向成脂、成骨分化.结论 单酶消化法是一种有效的分离培养UC-MSCs的方法,为建立MSCs库和临床应用提供了理论依据.

Objective To explore a new approach to isolate the umbilical cord-mesenchymal stem cells （UC-MSCs） from a whole umbilical cord. Methods Single enzyme was used to digest the whole umbilical cord,and passaged and cultured,draw the growth curve,cell cycle and cell wall antigen on the surface by flow cytometry. In particular inducing system, the adherent cells were induced into adipocytes and osteoblast. Results The mesenchymal stem cells （MSCs） were derived from the whole umbilical cord. The growth curve showed that the cell doubling time was （24.15 ± 0.49） h and the percentage of G1phase cells was 82.66%.The adherent cells expressed the CD93,CD105,CD44,CD29and CD73, and did not express the CD34, CD45, CD31and human leukocyte antigen DR （HLA-DR）. UC-MSCs could differentiate into adipocytes and osteoblast. Conclusions Single enzyme approach is a good method to obtain UC-MSCs from whole human umbilical cord,and it provides a theoretical basis for the establishment of MSCs bank and clinical application.