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造血祖细胞是生成血细胞的原始细胞。它在数量和性质上的改变,都会影响血细胞的正常生成速率和功能。造血祖细胞的培养技术成为当前实验血液学和临床血液学研究中最活跃的领域。因目前造血祖细胞的培养方法尚未统一,所用的培养液、血清、刺激因子亦各不相同。因此就国内常用的方法结合其临床应用作一简要介绍。动物造血祖细胞培养技术一、多向性造血祖细胞 CFU-S 测定1.酒精擦净超净工作台,开紫外线灯30分钟。2.配20%1640马血清+青、链霉素。3.供体小鼠给药(动物造型)。4.受体小鼠致死量照射(850rad)。5.供体小鼠细胞悬液的制备:正常小鼠颈部脱臼处死后,剪开皮肤,取出股骨,用酒精纱布剥离肌肉,用培养液从股骨中冲出细胞,让细胞通过4(1/2)号针头的注射器,使成为单细胞悬液。6.有核细胞计数(种的细胞数为5×10~4)。7.配体系液:(1)细胞液为 x;(2)20%1640马血清为10x。
Hematopoietic progenitor cells are primitive cells that produce blood cells. Its quantitative and qualitative changes affect the normal rate and function of blood cells. The culture technology of hematopoietic progenitor cells has become the most active field in the current experimental hematology and clinical hematology research. Due to the current methods of culture of hematopoietic progenitor cells have not been unified, the culture medium used, serum, stimulating factors are also different. Therefore, the methods commonly used in China combined with its clinical application for a brief introduction. Animal hematopoietic progenitor cell culture technology First, the multi-directional hematopoietic progenitor cells CFU-S Determination 1. Alcohol clean clean bench, open the UV lamp for 30 minutes. 2. With 20% 1640 horse serum + cyan, streptomycin. 3. donor mice (animal modeling). Recipient mice lethal dose (850rad). 5. Preparation of donor mouse cell suspension: After normal mouse cervical dislocation was sacrificed, the skin was cut off, the femur was removed, the muscle was stripped with alcohol gauze, the cells were flushed out of the femur with the culture medium, and the cells were allowed to pass through the 4 (1 / 2) needles to make a single cell suspension. 6. Nucleated cell count (the number of cells is 5 × 10 ~ 4). 7. Ligand system solution: (1) cell solution is x; (2) 20% 1640 horse serum is 10x.