原核表达重组人白介素-3及其对脐带血造血干细胞体外增殖的影响

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目的研究原核表达重组人白介素-3(rhIL-3)及其对脐带血造血干细胞体外增殖的影响。方法构建pET-43.1-rhIL-3表达载体,克隆至BL21菌株,并进行中试发酵生产,所得包涵体经透析复性后,采用阳离子交换柱纯化,纯化产物送N-端测序。Ficoll密度梯度离心结合免疫磁珠法富集人脐带血中CD34+细胞,以不同浓度rhlL-3及干细胞因子(SCF)、Fms样酪氨酸激酶受体-3配体(FL)和促血小板生成素(TPO)因子组合培养1周,测定细胞总数,计算CD34+细胞增殖倍数。结果在SCF和FL存在下,rh
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