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目的 探讨在HGF、FGF_4诱导下骨髓单个核细胞能否向肝细胞分化。方法 找健康忐愿者,年龄4～50岁,胸骨抽取骨髓,用淋巴分离液分离出骨髓单个核细胞,用含2％胎牛血清和1×ITS的DMEM培养,分别用单独HGF、单独FGF_4、HGF和FGF_4联合刺激、无生长因子4种处理因素进行诱导培养,分别于诱导培养0、7、14、21、28天时,留取细胞,用免疫细胞化学法检测CK18、AFP、白蛋白,用PAS法进行糖元染色试验以验证诱导分化的结果。结果 0天时,AFP、白蛋白少数细胞阴性,CK18阴性,糖元未见阳性;加生长因子的三组AFP在7、14天时表达逐渐增高,21、28天减弱;糖元、CK18、白蛋白表达逐渐增高。无生长因子组,在7、14、21、28天时,这此指标均未见表达。结论 在HGF、FGR_4诱导下,骨髓单个核细胞可分化为具肝细胞表型和功能的细胞。
Objective To investigate whether bone marrow mononuclear cells can differentiate into hepatocytes induced by HGF and FGF_4. Methods To find healthy volunteers, aged 4 to 50 years old, bone marrow was collected from the sternum, bone marrow mononuclear cells were isolated by lymphatic drainage and cultured in DMEM containing 2% fetal bovine serum and 1 × ITS. HGF, FGF4 alone , HGF and FGF_4 were stimulated with 4 kinds of factors without induction of growth factors were cultured on induction 0,7,14,21,28 days, cells were collected, immunocytochemical detection of CK18, AFP, albumin, Glycogen staining was performed using the PAS method to verify the result of induced differentiation. Results At 0 days, AFP, a few albumin cells were negative and CK18 was negative, but no glycogen was found in the three groups. The expression of AFP in the three groups increased gradually at 7 and 14 days and weakened at 21 and 28 days. Glycogen, CK18 and albumin Expression gradually increased. No growth factor group, at 7,14,21,28 days, this index were not expressed. Conclusion Bone marrow mononuclear cells can differentiate into cells with hepatocyte phenotype and function under the induction of HGF and FGR_4.