人脐带间充质干细胞的分离、鉴定及向肝细胞的定向诱导分化

Study on isolation and identification of mesenchymal stem cells from human umbilical cord and its hepatic differentiation

目的 分离纯化人脐带间充质干细胞（UCMSCs）,探讨UCMSCs是否能在体外条件下被诱导成为肝细胞样细胞.方法 无菌条件下采集健康新生儿脐带,采用胰酶、胶原酶顺序消化法分离单个细胞,通过贴壁培养方法分离纯化间充质干细胞（MSCs）.从形态、表面抗原、成脂肪细胞和成骨细胞分化潜能三方面进行鉴定.随后通过细胞因子诱导的方法将间充质干细胞诱导为肝细胞样细胞.结果 从人脐带中成功分离纯化得到UCMSCs,细胞呈纤维状;表达CD44、CD105、CD73、CD90,不表达与造血细胞和内皮细胞相关的CD34、CD45、CD31等标志物,不表达与移植物抗宿主病相关的HLA-DR等分子;在不同细胞因子作用下,可以被诱导分化为脂肪细胞、成骨细胞和卵圆形肝细胞样细胞.结论 人脐带中含有较丰富的MSCs,在体外条件下可以被诱导为肝细胞样细胞.

Objective To isolate and purify the mesenchymal stem cells from human umbilical cord（UCMSCs）,and to study their ability to differentiate mto hepatocyte in vitro under the induction of cytokines. Methods UCMSCs based on the two steps digestion were isolated and purified. MSCs were purified by centrifuge and attachment and identified from morphology, surface markers and differentiation potential, and then MSCs were differentiated into hepatocyte in vitro under the induction of cytokines. Results UCMSCs expressed CD44, CD105, CD73 and CD90, but didn＇t express antigens of haemopoietic such as CD34, CD45 and antigens related to graft versus host disease such as HLA-DR.UCMSCs could be differentiated into adipocytes, osteoblasts and oval hepatic-like cells. Conclusions UCMSCs have multiple differentiation potency, which could differentiate into cells with features of hepatocytes.