Zhai Qiong-li, Qiu Lu-gui, Liu Yan, Li Qian, Han Jun-ling, Zhou Zheng, Li Xin, Ying Hong-guang, Han Zhong-chao
State Key Lab of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, CAMS, PUMC, Tianjin 300020, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2002 Feb;24(1):7-10.
To compare the expression of cell adhesion molecules (CAMs) among VLA-4 (CD49 d), VLA-5 (CD49e), LFA-1 (CD11a), L-selectin (CD62L), and PECAM-1 (CD31) which are more related to the homing of hematopoietic stem and progenitor cells (HSPC) on the ex vivo expanded CD34+ subset with that of fresh isolated AC133+ cells.
AC133+ cells selected from fresh cord blood (CB) samples were cultured in QBSF-60 serum-free media in the presence of Flt-3 ligand + SCF + TPO (FST), with initial addition of IL-3 for up to 2 week. Expansion potential and the expression of above CAMs were evaluated at day 0, day 7, day 10 and day 14.
(1) Simultaneously numerical expansion of various HSPC was constantly observed during the culture, and the fold expansion of AC133+ cells and CD34+ cells on day 14 were 33.50 and 64.56 respectively; (2) The number of CD34+ subsets expressing the above adhesions were all increased at different degrees (from 20 fold to 160 fold). (3) The expressions of CD11a, CD49d, and CD49e on ex vivo expanded CD34+ cells were increased as compared to their baseline levels, but the percentage of CD62L+ and CD31+ subpopulations in CD34+ cells were decreased.
Our short-term culture system can not merely support the simultaneous expansion of CB derived AC133+ cells, but the expanded hematopoietic progenitors may well sustained the expression of homing-related adhesion molecules.
比较与造血干细胞和祖细胞(HSPC)归巢更相关的细胞黏附分子(CAMs),即VLA-4(CD49d)、VLA-5(CD49e)、LFA-1(CD11a)、L-选择素(CD62L)和PECAM-1(CD31)在体外扩增的CD34+亚群与新鲜分离的AC133+细胞中的表达情况。
从新鲜脐带血(CB)样本中筛选出的AC133+细胞在含Flt-3配体+SCF+TPO(FST)的QBSF-60无血清培养基中培养,最初添加IL-3培养长达2周。在第0天、第7天、第10天和第14天评估扩增潜能及上述CAMs的表达。
(1)培养期间持续观察到各种HSPC同时出现数量扩增,第14天AC133+细胞和CD34+细胞的扩增倍数分别为33.50和64.56;(2)表达上述黏附分子的CD34+亚群数量均有不同程度增加(从20倍到160倍)。(3)与基线水平相比,体外扩增的CD34+细胞上CD11a、CD49d和CD49e的表达增加,但CD34+细胞中CD62L+和CD31+亚群的百分比降低。
我们的短期培养系统不仅能支持源自CB的AC133+细胞的同时扩增,而且扩增后的造血祖细胞可能很好地维持归巢相关黏附分子的表达。