摘要
目的 探讨骨髓基质细胞培养上清 (SN)协细胞因子对人脐血造血细胞体外生长的影响。方法 使用rIL 3,rI 1β ,rIL 6 ,和SCF等细胞因子或SN或者两者联用长期培养人脐血造血细胞 ,进行细胞增殖和表面标志测定。结果 培养 7d细胞已开始增殖 ,14d达高峰 ,细胞总数在细胞因子组 (1)增加了 134.5倍和SN组 (3)增加 81.5倍 ,两者联用 (2 )增加了 171.3倍 ,2 1d开始下降 ,(3)组下降最明显 ,35d时细胞总数为培养前的 (3)组 2 6 .6 5倍 ,(1)组 5 4 .15倍 ,(2 )仍组有 92 .2 5倍。CD34+ 细胞第 7天已升高 ,14d达高峰 ,(1)、(3)组和 (2 )组分别比培养前增加了 6 8.4 3%、6 9.77%和 82 .4 7% ,2 1d开始减少 ,分别比培养前增加 5 6 .16 %、12 .33%和 71.72 % ;CD33+ 细胞 7天时均显著增加 ,14d达高峰 ,分别比培养前增加了 80 .6 7%、82 .14 %和 6 9.6 9% ,2 1d时维持在 14d水平 ;CD38+细胞 7d时均有显著增加 ,分别比培养前增加了 6 6 .6 5 %、70 .4 2 %和 73.8% ,14～ 2 1d均在该水平维持 ,三组间无显著性差异。结论 骨髓基质细胞培养上清可促进脐血造血细胞的增殖 ,但比细胞因子作用弱 ,两者联用时作用明显强于两者分别使用 ,提示骨髓基质细胞还可产生其它物质刺激造血细胞增殖。
Objective To explore the effect of supernatant (SN) from cultured human marrow stromal cells synergistically with cytokines on grouth of human umbilical cord blood (UCB) hemapoietic cells in vitro.Methods Hemapoietic cells from human UCB were cultured with cytokines( including rIL-3,rIL-6,rIL-1β and SCF) or stromal SN, or both of them together in a long term culture system. Their proliferation and change of cell suface markers were observed.Results Hemapoietic-4- cells proliferated at the 7th day and reached the highest level at the 14th day. The total nucleated cells increased 134.5-fold in the cytokines group(1), 81.5-fold in the SN group(3) and 171.3-fold in the group of both of them(2),then decreased gradually at the 21th day, and only increased 54.15-fold,26.65-fold and 92.25-fold respectively at the 35th day. CD34 + cell increased at the 7th day in every group, and reached the highest level at the 14th day,the percentage of increased 68.43% in (1)group、69.77% in (3)group and 82.47% in (2)group respectively,then decreased gradually and only increased 56.16%、12.33% and 71.72% than at starting culture respectively at the 21th day.CD33 + cell increased markedly at the 7th day in every group , and reached the highest level and increased 80.67% in(1)group、82.14% in (3)group and 69.69% in(2)group respectively at the 14th day, then maintained at this level at the 21th day.CD38 + cell increased markedly at the 7th day in every group, increased 66.65% in(1)group、70.42% in (3)group and 73.8% in(2)group respectively, then maintained at this level in every group at the 14th～21th day.Conclusion The supernatant derived from stromal cell culture promotes proliferation of hemapoietic cell from human UCB, but is inferior to cytokines. When both of them were used together, this role is obviously stronger than they were used alone. It shows that marrow stromal cells can produce other nuknowed factors to stimulate hemapoietic cell proliferation and prolong then culture in vitro.
出处
《江西医学院学报》
2004年第5期9-12,共4页
Acta Academiae Medicinae Jiangxi
关键词
骨髓基质细胞
上清
脐血
造血细胞
体外扩增表面标志
marrow stromal cells
supernatant
umbilical cord blood
hemapoietic cells
expand in vitro
suface markers
