Conversion of mononuclear cells from human umbilical cord blood into hepatocyte-like cells

Objective: To evaluate the differentiation of human umbilical cord blood cells into hepatocyte-like cells. Methods: Mononuclear cells (MNCs) derived from human umbilical cord blood were isolated using Ficoll. The experiment was derived into 3 categories:(1) MNCs co-cultured with 50 mg minced liver tissue separated by a trans-well membrane and then collected at 0 h,24 h,48 h and 72 h; (2) MNCs cultured along supplemented with 100 ml/L FBS. 100μ/ml penicillin, 100μg/ml streptomycin. 4. 7μg/ml linoleic acid, 1×ITS, 10-4 mol/L L-Ascorbic acid 2-P and a combination of FGF4 (100 ng/ml) and HGF (20 ng/mL). Cells were then collected at 0 d and 16 d to examine the expression profile of hepatocyte correlating markers;(3) 0. 2-0. 3 ml of MNCs with a cell density of 2×107/ml were transplanted into prepared recipient mice [n = 12, injected with 0. 4 ml/kg (20%) CCl4 and 150 ng/kg 5-fluorouracil (5-Fu) prior the transplant 24 h and 48 h, respectively] via injection through tail vein. Mice were sacrificed 4 weeks after transplantation. The hepatocyte correlating mRNAs and proteins were determined by RT-PCR, immunohistochemical analysis and immunoflurence technique. Results: (1) After 72 h. a number of glycogen positive stained cells were observed with MNCs co-cultured with damaged mouse liver tissues. The expression of hepatocyte markers, human albumin (ALB),α-fetal protein (AFP) and human GATA4 mRNA and proteins were detected by RT-PCR and immunohistochemistry as well. For the confirmation, the DNA sequencing of PCR products was performed. In control groups, MNCs co-cultured with normal mouse hepatocytes or MNCs cultured alone, all markers remained negative. (2) In growth factor supplemented culture system, MNCs developed into larger volume with richer cytoplasm and binucleation after 16 d. Positive expression of ALB, AFP, CK18 and CK19 mRNA were detected with RT-PCR, and ALB positive staining was observed by immunocytochemistry as well. In contrast. MNCs cultured without exogenous growth factors scarcely attached to the culture dish and ALB mRNA was not detected. (3) In transplantation experiment, both of ALB and AFP mRNA were detected by RT-PCR and HSA, PCNA and ALB positive staining were observed in the livers of recipient mice by immunocytochemistry. Conclusion: MNCs from human umbilical cord blood could convert into hepatocyte-like cells in 3 different ways, indicating their potential use in the clinic applications for the treatment of human liver diseases.     

人脐血细胞移植对高血糖模型小鼠的治疗作用

目的 探讨尾静脉移植人脐血细胞对糖尿病模型小鼠血糖的影响.方法 选择60只昆明小白鼠,随机抽取5只作为对照组,其余55只采用腹腔注射链脲佐菌素(STZ)的方法制备糖尿病鼠模型.成模后将小鼠随机分成移植组(26只)和糖尿病组(12只).将分离的人脐血单个核细胞于对数增长期用PKH26标记后,以2×106/ml经尾静脉注射入移植组小鼠体内.于注射后第4天和第14天观察3组小鼠的血糖和胰岛素水平,并用流式细胞仪和荧光显微镜观察PKH26标记的人脐血单个核细胞在体内的分布.结果 人脐血单个核细胞植入第4天,移植组小鼠血糖和胰岛素水平与糖尿病组、对照组比较,差异均有统计学意义(P＜0.05);移植第14天移植组小鼠血糖水平及胰岛素水平与糖尿病组比较,差异均无统计学意义(P＞0.05).流式细胞检测发现移植第4天的糖尿病小鼠的胰、脾、肝、骨髓组织内均可见PKH26+细胞,而移植第14天上述各部位未发现PKH26+细胞,荧光显微镜观察上述部位的冷冻切片结果也支持这一结果.结论 将人脐血细胞移植到未用免疫抑制剂的糖尿病小鼠体内可存活,并可以在一定时间内降低糖尿病小鼠的血糖水平.     

关节突关节射频热凝治疗老年性胸腰椎压缩性骨折后疼痛效果评价

目的:评价关节突关节射频热凝治疗老年性胸腰椎压缩性骨折后的疼痛效果.方法:挑选在该院进行治疗的老年性胸腰椎压缩性骨折患者62例,患者的收治时间均在2012年8月至2015年3月,将62例老年性胸腰椎压缩性骨折患者随机分成两组,每组各有患者31例,其中一组患者接受常规治疗,称为常规组,另一组患者接受关节突关节射频热凝治疗,称为研究组,记录常规组患者及研究组患者在治疗前、治疗后1天、一周、一个月及三个月后的疼痛情况,进行比较.结果:常规组患者与研究组患者在治疗前疼痛程度无明显差异,不存在统计学意义(即P ＞0.05);在治疗后1d、1w、1个月及3个月,研究组患者的疼痛程度均明显低于常规组患者,差异存在统计学意义(即P＜0.05).结论:对老年性胸腰椎压缩性骨折患者进行关节突关节射频热凝治疗能有效改善患者的疼痛症状,疗效确切,推荐临床应用.