Ma Li-Jun, Gao Lei, Zhou Hong, Qiu Hui-Ying, Hu Xiao-Xia, Xie Lin-Na, Wang Jian-Min
Department of Hematology, Changhai Hospital, The Second Military medical University, Shanghai 200433, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Oct;14(5):949-54.
To investigate the effects of human mesenchymal stem cells (MSC) and human fibroblastoid cell line (HFCL) as feeder layer on expansion of umbilical cord blood CD34(+) cells in vitro, (60)Co gamma-ray irradiated MSC and HFCL were used as feeder layer to expand cord blood CD34(+) cells in culture. The efficiencies of MSC and HFCL on expansion of CD34(+) cells in culture with or without cytokines were compared. The results showed that no matter whether cytokines (rhFL, rhSCF, rhTPO) were added, the proliferation of nucleated cells after expansion for 12 days in HFCL group was statistically higher than that in MSC group, i.e. with cytokines (9797 +/- 361)% vs (7061 +/- 418)%; without cytokines (5305 +/- 354)% vs (1992 +/- 247)%, when the cell numbers at day 0 was accounted as 100%), P < 0.01. The proliferation of propagated CD34(+) cells between MSC group and HFCL without addition of cytokines was not statistically different (820 +/- 191)% vs (825 +/- 305)%, P > 0.05. However, in the presence of cytokines, the propagating rate of MSC group was lower than that of HFCL group (939 +/- 212)% vs (1617 +/- 222)%, P < 0.01. MSC was better than HFCL in maintaining the LTC-IC of UCB CD34(+) cells, i.e. the number of CFU-GM colonies in the fifth week was (129.95 +/- 8.73) /10(5) seeded cells vs (89.81 +/- 10.29) colonies/10(5) cells, P < 0.05; with addition of cytokines, the effect was more obvious, i.e. the number of CFU-GM colonies in the fifth week (192.93 +/- 4.95)/10(5) seeded cells vs (90.47 +/- 14.28) colonies/10(5) seeded cells, P < 0.01. MSC mixed with a certain proportion of HFCL facilitated maintaining the LTC-IC of UCB CD34(+) cells. When the proportion was 4:1, the number of CFU-GM colonies was the highest (186.89 +/- 11.11)/10(5) seeded cells, which was higher than that of both 3:2 group [(138.92 +/- 14.84) colonies/10(5) seeded cells] and MSC only group, i.e. (64.63 +/- 6.11) colonies/10(5) seeded cells, both P < 0.01. It is concluded that HFCL is better than MSC in maintaining the expansion of CD34(+) cells and cytokines can enhance this effect, while MSC are stronger than HFCL in maintaining the LTC-IC of UCB CD34(+) cells in vitro. MSC with addition of a certain proportion of HFCL can significantly enhance the efficiency of CD34(+) cell expansion.
为研究人间充质干细胞（MSC）和成纤维样细胞系（HFCL）作为饲养层对体外扩增脐带血CD34(+)细胞的影响，将经（60）Coγ射线照射的MSC和HFCL用作饲养层，在培养中扩增脐带血CD34(+)细胞。比较了MSC和HFCL在有或无细胞因子情况下对培养中CD34(+)细胞的扩增效率。结果显示，无论是否添加细胞因子（rhFL、rhSCF、rhTPO），HFCL组扩增12天后有核细胞的增殖在统计学上均高于MSC组，即添加细胞因子时（9797±361）%对（7061±418）%；不添加细胞因子时（5305±354）%对（1992±247）%（以第0天的细胞数计为100%），P<0.01。在不添加细胞因子时，MSC组和HFCL组中扩增的CD34(+)细胞的增殖无统计学差异（820±191）%对（825±305）%，P>0.05。然而，在有细胞因子存在时，MSC组的扩增率低于HFCL组（939±212）%对（1617±222）%，P<0.01。在维持脐血CD34(+)细胞的长期培养起始细胞（LTC-IC）方面，MSC优于HFCL，即第5周时CFU-GM集落数为（129.95±8.73）/10(5)接种细胞对（89.81±10.29）个集落/10(5)细胞，P<0.05；添加细胞因子时，效果更明显，即第5周时CFU-GM集落数为（192.93±4.95）/10(5)接种细胞对（90.47±14.28）个集落/10(5)接种细胞，P<0.01。一定比例的MSC与HFCL混合有助于维持脐血CD34(+)细胞的LTC-IC。当比例为4:1时，CFU-GM集落数最高（186.89±11.11）/10(5)接种细胞，高于3:2组[（138.92±14.84）个集落/10(5)接种细胞]和仅MSC组，即（64.63±6.11）个集落/10(5)接种细胞，P均<0.01。结论是，在维持CD34(+)细胞扩增方面HFCL优于MSC，细胞因子可增强此效应，而在体外维持脐血CD34(+)细胞的LTC-IC方面MSC强于HFCL。添加一定比例HFCL的MSC可显著提高CD34(+)细胞的扩增效率。
