Baek Eun Jung, Kim Han-Soo, Kim Sinyoung, Jin Honglien, Choi Tae-Yeal, Kim Hyun Ok
The Department of Laboratory Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Transfusion. 2008 Oct;48(10):2235-45. doi: 10.1111/j.1537-2995.2008.01828.x. Epub 2008 Jul 31.
There is no appropriate alternative source of red blood cells (RBCs) to relieve the worsening shortage of blood available for transfusion. Therefore, in vitro generation of clinically available RBCs from hematopoietic stem cells could be a promising new source to supplement the blood supply. However, there have been few studies about the generation of clinical-grade RBCs by coculture on human mesenchymal stem cells (MSCs) and various cytokine supplements, even though the production of pure RBCs requires coculture on stromal cells and proper cytokine supplements.
Umbilical cord blood (CB) CD34+ cells were cultured in serum-free medium supplemented with two cytokine sets of stem cell factor (SCF) plus interleukin-3 (IL-3) plus erythropoietin (EPO) and SCF plus IL-3 plus EPO plus thrombopoietin (TPO) plus Flt-3 for 1 week, followed by coculture upon MSCs derived from bone marrow (BM) or CB for 2 weeks.
Almost pure clinical-grade RBCs could be generated by coculturing with CB-MSCs but not BM-MSCs. Expansion fold and enucleation rate were significantly higher in coculture with CB-MSCs than BM-MSCs. Despite a 2.5-fold expansion of erythroblasts in the presence of TPO and Flt-3 for 8 days, the final RBC count was higher without TPO and Flt-3.
This study is the first report on generating clinical-grade RBCs by in vitro culture with human MSCs and compared effectiveness of several cytokines for RBC production. This provides a useful basis for future production of clinically available RBCs and a model of erythropoiesis that is analogous to the in vivo system.
目前尚无合适的替代红细胞（RBC）来源来缓解日益严重的输血用血短缺问题。因此，从造血干细胞体外生成临床可用的红细胞可能是补充血液供应的一个有前景的新来源。然而，关于通过在人间充质干细胞（MSC）上共培养及添加各种细胞因子来生成临床级红细胞的研究很少，尽管生成纯红细胞需要在基质细胞上共培养并添加适当的细胞因子。
脐血（CB）CD34 + 细胞在补充有两组细胞因子的无血清培养基中培养，一组为干细胞因子（SCF）加白细胞介素 - 3（IL - 3）加促红细胞生成素（EPO），另一组为SCF加IL - 3加EPO加血小板生成素（TPO）加Flt - 3，培养1周，随后在源自骨髓（BM）或CB的MSC上共培养2周。
与CB - MSC共培养可生成几乎纯的临床级红细胞，而与BM - MSC共培养则不能。与BM - MSC相比，与CB - MSC共培养时的扩增倍数和去核率显著更高。尽管在TPO和Flt - 3存在下成红细胞在8天内扩增了2.5倍，但最终红细胞计数在没有TPO和Flt - 3时更高。
本研究是关于通过与人间充质干细胞体外共培养生成临床级红细胞以及比较几种细胞因子对红细胞生成有效性的首次报道。这为未来临床可用红细胞的生产提供了有用的基础以及一个类似于体内系统的红细胞生成模型。
