硫酸乙酰肝素协同细胞因子对脐血造血细胞体外生长的作用

Synergic effect of heparan sulfate and cytokines on in vitro growth of human umbilical cord blood hematopoietic cells

目的 探讨硫酸乙酰肝素 (HS)协同细胞因子对脐血造血细胞体外生长的影响。方法 细胞因子 (rhIL 3,rhIL 1β ,rhIL 6和SCF)、骨髓基质细胞培养上清 (SN) +细胞因子、SN、HS +细胞因子 4组分别与脐血造血细胞培养 ,检测细胞增殖、细胞表面标志和造血祖细胞集落生长。结果 培养至第14天细胞增殖达高峰 ,4个组细胞总数依次增加了 134 .5 ,171.3,81.5和 16 7.2倍。第 2 1天细胞数量SN组明显下降。CD3 4+细胞数第 7天已升高 ,第 14天达高峰 ,依次增加了 6 8.4% ,82 .5 % ,6 9.8%和79 3% ,第 2 1天仍为 5 6 .2 % ,71.7% ,12 .3%和 73.7%。CD3 3+细胞数第 14天达高峰 ,依次增加了80 2 % ,6 8.6 % ,81.6 %和 70 .3% ,第 2 1天时仍维持在第 14天水平。CD3 8+细胞数第 7天时比培养前增加了 6 6 .6 % ,73.8% ,70 .4%和 71.9% ,第 14至第 2 1天均维持该水平。扩增 1周各组CFU GM产率均有提高 ,2周达最高 ,依次增加了 2 5 0 % ,2 79% ,2 17%和 2 73%。第 3周仍有 15 1% ,2 40 % ,145 %和2 31%。结论 SN与细胞因子联用对脐血造血细胞增殖的促进作用明显强于两者单用。

Objective To explore the synergic effect of heparan sulfate(HS) and cytokines on the growth of human umbilical cord blood (UCB) hematopoietic cells. Methods Hematopoietic cells from human UCB were cultured with ①cytokines(rhIL-3,rhIL-1β,rhIL-6 and SCF) or ②SN(supernatant from cultured human marrow stromal cells) and cytokines, or ③ SN, or ④ HS and cytokines. Cellular proliferation, CFU-GM yields and changes of cell immunophenotype were observed. Results Hematopoietic cell proliferation reached peak at the 14th day . The number of total nucleated cells increased 134.5-, 171.3-, 81.5- and 167.2-fold in ①,②,③ and ④groups, respectively, at the 21th day, the ③ group significantly decreased. CD 34 + cells increased at the 7th day and reached peak, at the 14th day with a percentage of 68.4%, 82.5%, 69.8% and 79.3% ,and at the 21th day 56.2%, 71.7.%, 12.3% and 73.3%, respectively. CD 33 + cells reached peak at the 14th day and increased by 80.2%, 68.6%, 81.6% and 70.3%, respectively, and remained these levels at the 21th day . CD 38 + cells increased by 66.6%, 73.8%, 70.4% and 71.9% at the 7th day and remained this level at the 14th and the 21th day. From the first week of culture, the percentage of CFU-GM increased in all of the four groups, at the second week of culture, it increased by 250%, 279%, 217% and 273%, and at the third week still increased by 151%, 240%, 145% and 231%, respectively. Conclusion The combination of SN and cytokines have a synergic effect in promoting proliferation of hematopoietic cell from human UCB.The synergic effect remained the same when SN was replaced by heparan sulfate .