一种新型生物反应器在造血干/祖细胞体外扩增中的应用

The Application of a New-type Bioreactor in the ex vivo Expansion of Hematopoietic Stem/Progenitor Cells

针对造血干/祖细胞体外扩增对培养环境的需求,结合静/动态培养的特点,开发了一种新型的生物反应器用于造血干/祖细胞的体外扩增。在该生物反应器内,采用SCF+TPO+Flt-3细胞因子组合,比较了静态和循环培养两种方式体外扩增脐血CD34+细胞的效果。培养7d后,总细胞分别扩增了(13.86±4.26)和(7.23±2.67)倍,显示静态培养有利于总细胞的扩增;CD34+细胞扩增倍数、培养物中CD34+细胞含量均相近,无显著性差异;而CD34+CD38-细胞扩增倍数以及培养物中CD34+CD38-细胞的百分含量分别为(1.82±0.58)和(3.90±0.85)倍以及(9.45±4.85)和(37.47±14.06)%,循环培养明显高于静态培养。可见,在该生物反应器内,采用静态和循环两种培养方式,均能实现造血干/祖细胞的体外扩增,但静态培养促使造血干细胞向定向祖细胞分化,而循环培养则更有利于早期造血干细胞的扩增。

Based on the requirement of culture conditions for hematopoietic stem and progenitor ceils （HSPCs） ex vivo expansion, we developed a new-type bioreactor by combining superiorities of static and stirred culture models. Stem cell factor（SCF）, thrombopoietin（TPO）, FLT-3 ligand（Flt-3） were used as the cytokines cocktails. The effects of the static and cyclic culture on the expansion characteristics of CD34^＋ selected ceils were compared in the new-type bioreactor. After 7 d cultures, the expansion of total ceils in the static culture was 13.86 ± 4.26 fold, higher than that in the cyclic culture （7.23 ± 2.67 fold）. The analysis of the fold expansion and the proportion of CD34^＋ ceils showed that there was no marked difference between the static culture and the cyclic culture. However, the fold expansion and the proportion of CD34^＋CD38^- ceils were higher in the cyclic culture than those in the static culture （3.90 ± 0.85 versus 1.82 ± 0.58）, which reflected more primary CD34^＋CD38^- ceils were obtained in the cyclic culture. The above results demonstrated that both the static culture and the cyclic culture could be used in ex vivo expansion of CD34^＋ ceils with the new-type bioreactor. In static culture hematopoietic stem ceils differentiated into progenitor ceils, whilst the cyclic culture favored the expansion of primary HSPCs.