肝脏细胞条件培养基诱导大鼠骨髓间质细胞分化为肝细胞的作用

The Differentiation of Bone Marrow Stromal Cells into Hepatocyte by Conditioned Medium of Liver Cells in Vitro

目的 :探讨肝脏细胞条件培养基诱导大鼠骨髓间质细胞分化为肝细胞的作用。方法 :从大鼠骨髓中分离纯化培养间质细胞 ,诱导前 2 4小时加 1μg/L碱性成纤维生长因子 (bFGF)入培养液中以促进细胞分裂 ,再以肝脏细胞条件培养基作诱导剂 ,分别在诱导培养 0、 7、 14、 2 1、 2 8天时 ,留取细胞 ,观察细胞形态的变化 ,并采用免疫细胞化学方法检测肝细胞功能标志物 (AFP、白蛋白、CK18)、用PAS法进行糖元染色试验 ,以验证诱导分化的结果。结果 :诱导后 3天间质细胞表现为肝细胞样 ,随着诱导时间的延长 ,肝细胞功能标志逐渐出现和成熟。AFP在 7、 14天时表达较高 ,2 1、 2 8天时表达显著减少 ;白蛋白、CK 18和糖元随着诱导时间的延长表达逐渐增多。结论 :肝脏细胞条件培养基能诱导骨髓间质细胞分化为肝细胞。

Objective:To study whether the differentiation of bone marrow stromal cells(BMSCs) into hepatocyte by conditioned medium of liver cells in vitro.Methods:The BMSCs from rats were isolated and cultured for 6 passages.The cells were pretreated for 24h in DMEM containing 20% FBS(v/v) and 1μg·L -1 bFGF.Then the medium was replaced with the condition culture medium of liver cells for induction.The morphological changes of the cells were observed.Cells were collected on day 7 th ,14 th ,28 th .CK18,AFP and albumin were tested by immunocytochemistry.Glycogen was tested by periodic acid schiff(PAS).Results:Morphological changes of the stromall cells were noted 3 days after induction.Hepatic phenotype and fuction were appearing progressively with the cultured time longer.The expression of AFP increased progressively on day 7 th ,14 th ,then decreased on day 21 th ,28 th .The expressions of albumin,CK18 and glycogen were stronger with the cultured time longer.Conclusion:The results show that the bone marrow stromal cells can be induced to differentiate into hepatocyte like by conditioned medium of liver cells in vitro.