不同年龄段慢性肝衰竭患者自体骨髓干细胞体外增殖能力观察

Observation of proliferative capacity of human bone marrow mesenchymal stem cells in patients with chronic liver failure in vitro

目的观察不同年龄段慢性肝衰竭患者自体骨髓间充质干细胞体外增殖能力的特点。方法在慢性肝衰竭患者30例和非肝病患者30例,各分3个组,即A组为20～35岁,B组为35～55岁和C组为>55岁。取骨髓血50毫升,采用密度梯度离心法分离出单个核细胞,接种于56.7cm2培养皿内,观察细胞生长情况,并在第7天计数集落形成单位;当贴壁细胞融合后进行传代,计算增至治疗所需细胞数量的传代时间和细胞增殖率。结果在慢性肝衰竭患者,A组为6例,B组为18例,C组为6例;在非肝病组,分别为7、12和11例;在肝衰竭患者A组、B组和C组每皿细胞集落单位分别为15.3±8.7×106、8.6±5.2×106和4.2±3.6×106,而在非肝病患者则分别为14.7±7.1×106、13.5±7.4×106和10.3±4.9×10(6P<0.05);第一代细胞倍增时间比较,肝衰竭患者A组、B组和C组分别为178.7±79.6h、264.1±102.3h和457.7±155.6h,而非肝病患者则分别为156.3±62.5h、192.7±88.6h和311.6±142.8h(P<0.05);第二代细胞倍增时间比较,肝衰竭患者也明显比非肝病患者延长。结论大于55岁的慢性肝衰竭患者自体骨髓干细胞体外增殖能力明显降低。在选择干细胞移植治疗时,宜选择其他来源的细胞。

Objective To observe the proliferative eapacity of human bone marrow mesenchymal stem eells （MSCs） in patients with chronic liver failure in vitro. Methods MSCs were isolated from human bone marrow from 30 patients with chronic liver failure and 30 patients with benign diseases. In each group,the patients were divided into group A（younger than 30 year old）,group B（aged from 31 to 55 year old）and group C（older than 55 years）. The cell numbers were determined at the end of every passage,and the capability of proliferation was calculated by colony-forming units and double times at the first and second passage. Results There were six in group A, eighteen in group B and six in group C in patients with chronic liver failure,and they were seven, twelve and eleven patients with benign diseases;the colony-forming units in group A,B and C in patients with liver failure were 15.3±8.7×10^6,8.6±5.2×10^6 and 4.2±3.6×10^6,respectively,while in patients with benign diseases, they were 14.7±7.1×10^6,13.5±7.4×10^6 and 10.3±4.9×10^6,respectively （P〈0.05）;the double times of first passage in group A,B and C in patients with liver failure were 178.7±79.6h,264.1_±102.3h and 457.7±155.6h,respectively, while they were 156.3±62.5h,192.7±88.6h and 311.6±142.8h, respectively （P〈0.05）;in second passage,the double times in patients with liver failure prolonged obviously too, as compared to those in patients with benign diseases. Conclusion The proliferation capability of MSCs decreases in patients with chronic liver failure,especially in elderly patients.