人脐带血间充质干细胞对脐带血中CD34+细胞扩增的影响

[Effects of human umbilical cord blood mesenchymal stem cells on the expansion of CD34+ cells from umbilical cord blood].

作者信息

Zhou Dun-hua, Huang Shao-liang, Zhang Xu-chao, Wei Jing, Wu Yan-feng, Huang Ke, Li Yang, Fang Jian-pei

机构信息

Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510120, China.

出版信息

Zhonghua Er Ke Za Zhi. 2005 Jul;43(7):494-8.

PMID:16083546

Abstract

OBJECTIVE

The previous studies indicated that mesenchymal stem cells (MSCs) either from umbilical cord blood (UCB) or from bone marrow (BM) had the same biological characteristics and the function of secreting hematopoietic growth factors (HGFs). The present study aimed to understand the effects of human UCB MSCs on the expansion of CD(34)(+) cells from UCB.

METHODS

Human UCB CD(34)(+) cells were incubated in the system containing UCB MSCs, HGFs and serum free medium. 2. The surface markers (CD(34)(+), CD(34)(+)CD(38)(-), CD(34)(+)CD(3)(+), CD(34)(+)CD(19)(+), CD(34)(+)CD(33)(+), CD(34)(+)CD(41a)(+)) on expanded UCB cells were examined by flow cytometry on the 6th and 12th days. 3. The expanded and unexpanded cells were cultured in semi-solid culturing system and checked for colony forming units of granulocyte and macrophage (CFU-GM), erythroid burst-forming unit (BFU-E), colony forming units of granulocyte- erythrocyte-megakaryocyte-macrophage (CFU-Mix) and colony forming units of high-proliferative potential (CFU-HPP).

RESULTS

The expansion folds of CD(34)(+)CD(38)(-) cells from UCB MSCs + HGFs groups on the 6th and 12th days were 159.43 and 436.68, respectively. Interestingly, the percentage of CD(34)(+)CD(38)(-) cells declined in HGFs group after expanding for 12 days, but it rose to 9.98% in the UCB MSCs + HGFs group. 2. Colony forming capacity of expanded UCB cells showed that the folds of CFU-Mix and CFU-HPP of UCB MSCs + HGFs group increased from day 6 to day 12, but the folds decreased in the HGFs group. 3. From day 0 to day 12, CD(34)(+)CD(33)(+) cells and CD(34)(+)CD(41a)(+) cells were amplified gradually, but CD(34)(+)CD(19)(+) and CD(34)(+)CD(3)(+) cells decreased gradually, and in UCB MSCs + HGFs group this phenomenon was more significant than that in HGFs group.

CONCLUSION

UCB MSCs containing system not only has the ability to expand the primitive HSCs but also has the ability to sustain the proliferation of HSCs. 2. UCB MSCs containing system amplified mainly myeloid and megakaryocytoid progenitor subsets. These may have clinical significance in reducing infection and hemorrhage.

摘要

目的

先前的研究表明，来自脐带血（UCB）或骨髓（BM）的间充质干细胞（MSC）具有相同的生物学特性以及分泌造血生长因子（HGF）的功能。本研究旨在了解人脐带血间充质干细胞对脐带血中CD(34)(+)细胞扩增的影响。

方法

将人脐带血CD(34)(+)细胞置于含有脐带血间充质干细胞、造血生长因子和无血清培养基的体系中培养。2. 在第6天和第12天，通过流式细胞术检测扩增后脐带血细胞的表面标志物（CD(34)(+)、CD(34)(+)CD(38)(-)、CD(34)(+)CD(3)(+)、CD(34)(+)CD(19)(+)、CD(34)(+)CD(33)(+)、CD(34)(+)CD(41a)(+)）。3. 将扩增和未扩增的细胞在半固体培养体系中培养，检测粒细胞和巨噬细胞集落形成单位（CFU-GM）、红系爆式集落形成单位（BFU-E）、粒细胞-红细胞-巨核细胞-巨噬细胞集落形成单位（CFU-Mix）和高增殖潜能集落形成单位（CFU-HPP）。

结果

脐带血间充质干细胞+HGFs组中，第6天和第12天CD(34)(+)CD(38)(-)细胞的扩增倍数分别为159.43和436.68。有趣的是，HGFs组中CD(34)(+)CD(38)(-)细胞在扩增12天后百分比下降，但在脐带血间充质干细胞+HGFs组中升至9.98%。2. 扩增后脐带血细胞的集落形成能力显示，脐带血间充质干细胞+HGFs组的CFU-Mix和CFU-HPP倍数从第6天到第12天增加，但HGFs组的倍数下降。3. 从第0天到第12天，CD(34)(+)CD(33)(+)细胞和CD(34)(+)CD(41a)(+)细胞逐渐扩增，但CD(34)(+)CD(19)(+)和CD(34)(+)CD(3)(+)细胞逐渐减少，且在脐带血间充质干细胞+HGFs组中这种现象比HGFs组更显著。