Nakahata T, Ogawa M
J Clin Invest. 1982 Dec;70(6):1324-8. doi: 10.1172/jci110734.
We report identification of a unique class of human hemopoietic colony-forming cells with extensive ability to generate progenitors for secondary colonies. Mononuclear cells isolated from human umbilical cord blood formed colonies consisting of 40-500 blast cells after 25 d of incubation in methylcellulose culture in the presence of erythropoietin and medium conditioned by phytohemagglutinin-stimulated leukocytes. Replating of these blast cell colonies revealed that 100% of the primary colonies had the ability to generate secondary colonies, including multipotential colonies. These colonies could be distinguished from other hemopoietic colonies in situ by the complete absence of signs of terminal differentiation. Replating of granulocyte-erythrocyte-macrophage-megakaryocyte (GEMM) colonies, consisting of an average of 2 x 10(4) cells, revealed less capacity for secondary colony formation. This human blast cell colony assay may provide a method for quantitation of more primitive hemopoietic stem cells than progenitors for GEMM colonies (CFU-GEMM) in man.
我们报告鉴定出一类独特的人类造血集落形成细胞,其具有广泛的能力来产生次级集落的祖细胞。从人脐带血中分离出的单核细胞,在含有促红细胞生成素和经植物血凝素刺激的白细胞条件培养基的甲基纤维素培养基中培养25天后,形成了由40 - 500个原始细胞组成的集落。对这些原始细胞集落进行再培养发现,100%的初级集落有能力产生次级集落,包括多能集落。这些集落在原位可通过完全没有终末分化迹象与其他造血集落区分开来。对平均由2×10⁴个细胞组成的粒细胞 - 红细胞 - 巨噬细胞 - 巨核细胞(GEMM)集落进行再培养,结果显示其形成次级集落的能力较低。这种人类原始细胞集落测定法可能提供一种方法,用于定量比人类GEMM集落(CFU - GEMM)祖细胞更原始的造血干细胞。