四联袋分离脐血干细胞方法及临床效果初步研究

目的探讨一种高效实用、安全且具有临床应用价值的分离脐血干细胞方法，并研究多次、多份脐带血干细胞静脉叠加输注治疗股骨头坏死的临床安全性和疗效。方法①应用四联袋结合无菌接口技术制备脐血干细胞，对分离后单个核细胞（MNC）数、MNC收率、CD^34＋细胞数、台盼蓝拒染率、红细胞混入量进行检测。②将四联袋分离的脐血干细胞静脉输注给8例股骨头坏死患者，观察患者髋关节疼痛程度、性质及时间变化、髋关节外展与内旋功能变化、行走距离及步态变化；细胞移植后6个月，观察X线图像、股骨头形态学变化。结果①四联袋结合无菌接口技术分离的脐血MNC收率高，效果好且分离过程不易发生微生物污染。②8例患者临床症状均有不同程度的缓解或消失，有效率100％；细胞移植后6个月，X线图像、股骨头形态学有不同程度改善。③在细胞移植过程中及移植后均未发生并发症和发热、过敏等不良反应。结论①四联袋分离脐血干细胞是一种效率高、安全且具有临床应用价值的分离方法。②经静脉多次、多份输注脐血间质干细胞，是全身整体治疗股骨头坏死的一种新手段。③多份脐血干细胞不需ABO、HLA配型输注。

Method of Separating Stem Cells from Umbilical Cord Blood by Four Linked Bags and its Clinical Effects

[ Objective] To discuss a method of separating stem cells from human umbilical cord blood, which is efficient, practical and safe, as well as with clinical value. And to investigate the clinical safety and efficacy of multiple umbilical cord blood stem cells intravenous infusion to cure femoral head necrosis. [ Methods] ①Aseptic interface technology combined with four linked bags were adopt to produce umbilical cord blood stem cells. After separating, the number of mononuclear cells （ MNC ）and CD^34＋ cells, the yield rates of MNC, the exclusion rates oftrypan blue dye, the mixture of RBC were tested respectively. ②The umbilical cord blood stem cells separated by four linked bags was infused intravenously into 8 patients with femoral head necrosis. Which efficacy were obServed, including hip pain, pain characters, pain time, the nature of changes in the hip outreach and internal rotation function, the walking distance and gait changes. After 6 months of cell transplantation, X-ray images and morphological changes of the femoral head have been observed. [ Results] ①The method of separating stem cells from umbilical cord blood by four linked bags combined with aseptic interface technology was effective, and had high yield rate of umbilical cord blood MNC. Furthermore, the microbial contamination hardly occurred in the process of separation. ②The clinical symptoms were eased or disappeared in different levels in 8 cases with the effective rates of 100%. After 6 months of cells transplantation, X-ray images and the femoral head morphology have been improved in various degrees. （~） Complications, fever,, allergies or other adverse reactions have not happened in the process or after transplantation. [ Conclusion ] ①The method of separating umbilical cord blood tern cells by four linked bags is effective, safe and with clinical values. ②Multiple umbilical cord blood-derlved mesenchymal stem cells transplantation with intravenous infusion is a new holistic approach for femoral head necrosis. ③Sever