Tian Hong, Zheng Jin-e, Gong Fei-li, Wang Xing-bing, Huang Shi-ang, Chen Zhong
Department of Biology and Medical Genetics, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430030, China.
Zhonghua Xue Ye Xue Za Zhi. 2005 May;26(5):257-60.
To cultivate hematopoietic stem/progenitor cells (CD34(+)CD38(-)) isolated from umbilical cord blood (UCB) long for the observation of cell growth and expansion in vitro, surface marker expression, and chromosomal complements.
By flow cytometry CD34-FITC and CD38-PE labeled CD34(+) and CD38(-) stem/progenitor cells were isolated from UCB. The cells were cultivated in vitro for 6 months in a stem cell culture system with addition of six kinds of cell growth factors (IL-3, IL-6, GM-CSF, Epo, SCF, IGF-1). One month after cultivation, cultured cells were investigated for surface marker expression by flow cytometry and karyotype by G banding method.
After 7-12 days cultivation, the CD34(+)CD38(-) stem/progenitor cells began proliferation. The proliferation rate and the peak proliferation duration were greater in 1 cell/well cultivation conditions than in 10 cells/well. The cells remained CD34(+)CD38(-) and their karyotypic characteristics remained unchanged.
CD34(+)CD38(-) stem/progenitor cells from UCB may provide a larger than original amount of stem/progenitor cells for transplantation after long-term cultivation in vitro.
培养从脐带血（UCB）中分离出的造血干/祖细胞（CD34(+)CD38(-)），用于观察其体外细胞生长与扩增、表面标志物表达及染色体组成。
通过流式细胞术，利用CD34-FITC和CD38-PE标记从脐带血中分离出CD34(+)和CD38(-)干/祖细胞。将细胞在添加六种细胞生长因子（IL-3、IL-6、GM-CSF、Epo、SCF、IGF-1）的干细胞培养体系中体外培养6个月。培养1个月后，通过流式细胞术检测培养细胞的表面标志物表达，并采用G显带法检测核型。
培养7 - 12天后，CD34(+)CD38(-)干/祖细胞开始增殖。在1个细胞/孔的培养条件下，增殖速率和增殖高峰持续时间均高于10个细胞/孔的培养条件。细胞仍保持CD34(+)CD38(-)，其核型特征未发生改变。
脐带血来源的CD34(+)CD38(-)干/祖细胞经体外长期培养后，可为移植提供比原来数量更多的干/祖细胞。
