Ostasiewicz L T, Fleischman J, Poh-Fitzpatrick M B
Department of Dermatology, New York Medical College, Valhalla 10595.
Anal Biochem. 1993 Sep;213(2):395-9. doi: 10.1006/abio.1993.1437.
A simple, reproducible, and economical procedure for separating bone marrow cells into distinct populations and stages of maturation was developed and used to investigate the distribution of ferrochelatase activity in rat bone marrow cells. Density gradient ultracentrifugation of normal rat marrows on discontinuous arabinogalactan layers of 16.5-30.0% resulted in six distinct fractions enriched for myeloblasts, neutrophils, pronormoblasts, normoblasts, or lymphocytes. Segregation reflecting different stages of cell development was demonstrated. Neutrophils appeared at a greater density and were clearly separated from precursor myeloblasts. Normoblasts appeared at a greater density and were clearly separated from precursor pronormoblasts. Lymphocyte distribution within the gradient followed a normal curve (P < 0.05). A single gradient layer which consisted of greater than 80% lymphocytes was shown to be associated with significantly increased (P < 0.05) ferrochelatase specific activity (units/mg protein); the remaining cell populations did not differ significantly from each other in specific activity.
开发了一种简单、可重复且经济的方法,用于将骨髓细胞分离成不同的群体和成熟阶段,并用于研究大鼠骨髓细胞中铁螯合酶活性的分布。在16.5 - 30.0%的不连续阿拉伯半乳聚糖层上对正常大鼠骨髓进行密度梯度超速离心,得到了富含成髓细胞、中性粒细胞、早幼红细胞、正成红细胞或淋巴细胞的六个不同组分。证明了反映细胞发育不同阶段的分离情况。中性粒细胞出现在更高的密度处,并与前体成髓细胞明显分离。正成红细胞出现在更高的密度处,并与前体早幼红细胞明显分离。梯度内淋巴细胞的分布呈正态曲线(P < 0.05)。显示由大于80%淋巴细胞组成的单个梯度层与铁螯合酶比活性(单位/毫克蛋白质)显著增加(P < 0.05)相关;其余细胞群体在比活性上彼此无显著差异。