Sitar G
Br J Haematol. 1984 Nov;58(3):465-72. doi: 10.1111/j.1365-2141.1984.tb03993.x.
This paper reports a simple procedure for obtaining human megakaryocytes with a high purification and high recovery yield. Bone marrow cells, obtained from surgically removed ribs, were separated by a two-step procedure. Initially, a single cell suspension was enriched in megakaryocytes by equilibrium density centrifugation, the low density cell fraction was subsequently layered over a shallow continuous albumin gradient in a glass sedimentation chamber. Megakaryocytes averaged 0.03 +/- 0.02% of all nucleated cells in the starting marrow cell suspension, after this procedure an average 80 +/- 15% of the initial megakaryocyte population was recovered with a purity of 94 +/- 4%. Previous methods, based upon the use of a two-step procedure, are reviewed. The theory of velocity sedimentation is discussed with regard to the differences in the methodology used, which account for the different results I obtained.
本文报道了一种获得高纯度、高回收率人巨核细胞的简单方法。从手术切除的肋骨中获取的骨髓细胞，通过两步法进行分离。首先，通过平衡密度离心法富集巨核细胞形成单细胞悬液，随后将低密度细胞组分铺在玻璃沉降室中的浅连续白蛋白梯度上。在起始骨髓细胞悬液中，巨核细胞平均占所有有核细胞的0.03±0.02%，经过此步骤后，平均回收了初始巨核细胞群体的80±15%，纯度为94±4%。回顾了以前基于两步法的方法。讨论了速度沉降理论与所用方法差异的关系，这些差异解释了我所获得的不同结果。
