脐带血造血干／祖细胞体外扩增制剂及其临床前安全性检测分析  

Analysis on expansion potentials and pre-clinical safeties of hematopoietic stem cells from human umbilical cord blood.

作　　者：郭春娟[1] 侯迪[1] 高缨[2] 席咏梅[1] 王金福[1] 

机构地区：[1]浙江大学细胞与遗传学研究所,浙江杭州310058 [2]杭州市第二人民医院妇产科,浙江杭州310015

出　　处：《浙江大学学报（理学版）》2009年第3期328-334,共7页Journal of Zhejiang University（Science Edition）

摘　　要：造血干细胞移植(HSCT)可用来治疗多种恶性造血系统疾病.HSCT成功植入的先决条件之一是必须输入足够量的造血干/祖细胞(HS/PCs).细胞剂量过少会导致植入时间延长,移植失败的可能性增大.从脐带血中直接分离的造血干细胞因数量较少而不能满足临床移植的需要.利用骨髓间充质干细胞作为滋养层联合细胞因子组合(SCF+G-CSF+TPO)使脐血造血干/祖细胞体外扩增75.2±15.0倍,CD34+细胞扩增33.4±12.3倍.将扩增的细胞收集,重新悬浮于L-15培养液中制成造血干/祖细胞制剂,并在建立相应的检测技术基础上,进行了细胞制剂的无菌检测以及细胞制剂中细菌内毒素、牛血清残留量和干细胞因子残留量的检测.通过上述检测分析,为扩增的造血干/祖细胞制剂服务临床需要,辅助治疗恶性血液病和再生障碍性贫血以及遗传性贫血重建造血功能提供实验依据.Hematopoietic stem cell transplantation （HSCT） was widely used in treatment of hematologic malignancies. Sufficient hematopoietic/progenitor stem cells （HS/PCs） were critical for successful transplantation. A low cell dose would results in a delayed time to engraftment and high rates of engraftment failure. Insufficiency of cells is a main obstacle in clinical transplantation. When it is co-cultured with bone marrow-derived mesenchymal stem cells （BM-MSC） in the presence of cytokines, the cell growth was significantly enhanced： total cells by 75.2±15.0 fold, CD34^＋ cells by 33. 4±12. 3-fold. The expanded cells were collected and prepared as the expansion product of HS/PCs by resuspending cells in Leibovitz＇s L-15 Medium. Then, we established a series of methods to determine the contamination of bacteria, fungi and mycoplasmas, the bacterial endotoxin concentration, the residual bovine serum and SCF contents.

关 键 词：造血干/祖细胞 共培养 体外扩增 安全性 

分 类 号：Q297[生物学—细胞生物学]