Kelly G, Gasson J, Robinson M, Eiseman B
Surgery. 1977 Aug;82(2):260-5.
The long-range objective of this study is in vitro tissue culture of bone marrow stem cells to produce erythroid cells of sufficient volume for clinical transfusion. Bone marrow from dogs and patients was cultured in 29 experiments lasting up to 15 weeks. Peripheral erythroid cells from dogs were cultured in three control experiments. Optimal tissue culture media was NCTC 109 augmented with vitamin B12, erythropoietin (EP), folic acid, and 9% fetal protein in 60 mm glass Petri dishes. Additional media and Step III EP was added at 2 to 4 day intervals. Peripheral erythroid cells in culture all were dead within 4 weeks. Marrow erythroic cells in culture proliferated as demonstrated by (1) Fe59 incorporation into cells during culture, (2) H3 thymidine uptake into cultured cells, (3) microscopic evidence of mitoses, and (4) total erythrocyte concentration in cultures far exceeding that of peripheral culture controls. For as yet unexplained reasons the total mature red blood cell concentration in the culture media remained essentially constant throughout the studies. This is a first step in achieving the ultimate goal of bulk erythrocyte production from tissue culture.
本研究的长期目标是对骨髓干细胞进行体外组织培养，以生产出足够数量的红细胞用于临床输血。在29项持续长达15周的实验中，对狗和患者的骨髓进行了培养。在三项对照实验中，对狗的外周红细胞进行了培养。最佳组织培养基是在60毫米玻璃培养皿中添加了维生素B12、促红细胞生成素（EP）、叶酸和9%胎蛋白的NCTC 109。每隔2至4天添加额外的培养基和第三步EP。培养中的外周红细胞在4周内全部死亡。培养中的骨髓红细胞增殖，表现为：（1）培养期间Fe59掺入细胞；（2）H3胸腺嘧啶核苷摄入培养细胞；（3）有丝分裂的显微镜证据；（4）培养物中的总红细胞浓度远远超过外周培养对照。由于尚未明确的原因，在整个研究过程中，培养基中成熟红细胞的总浓度基本保持不变。这是朝着通过组织培养大量生产红细胞这一最终目标迈出的第一步。
