Jehn U, Oerkermann H, Heller A, Hötzel J
Blut. 1976 Oct;33(4):235-45. doi: 10.1007/BF00995219.
Bone marrow from 52 healthy donors has been fractionated on two discontinuous albumin gradients with different osmolarities in an effort to eliminate immunocompetent cells and to isolate hemopoietic stem cells. The nutrient agar system of Robinson was utilised as an assay procedure for the detection of colony-forming cells (CFU-c) in the resulting fractions. CFU-c, early granulocytic progenitors, are consistently present in the lighter fractions. On cytochemical staining, they showed a positive reaction for enzymes bound to lysosomes. The concentration of CFU-c was in an average 7 times that of the original marrow. At best, 1 X 10(5) CFU-c could be isolated from 10 ml marrow corresponding to 2 X 10(8) buffy-coat cells. The presence of immunocompetent cells was monitored by Phytohemagglutinin (PHA)-stimulation and T-cell rosette formation. In a human marrow transplant situation it appears that a better yield of CFU-c is necessary than is afforded by albumin gradient technique.
为了清除免疫活性细胞并分离造血干细胞,我们用两种不同渗透压的不连续白蛋白梯度对52位健康供者的骨髓进行了分级分离。采用罗宾逊的营养琼脂系统作为检测方法,以检测所得分级分离物中的集落形成细胞(CFU-c)。CFU-c,即早期粒细胞祖细胞,始终存在于较轻的分级分离物中。经细胞化学染色,它们对与溶酶体结合的酶呈阳性反应。CFU-c的浓度平均为原始骨髓的7倍。最多可从10毫升骨髓(相当于2×10⁸个血沉棕黄层细胞)中分离出1×10⁵个CFU-c。通过植物血凝素(PHA)刺激和T细胞玫瑰花结形成来监测免疫活性细胞的存在。在人类骨髓移植情况下,似乎需要比白蛋白梯度技术获得更高产量的CFU-c。