Carow C E, Hangoc G, Cooper S H, Williams D E, Broxmeyer H E
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202-5121.
Blood. 1991 Nov 1;78(9):2216-21.
The replating capability of human multipotential (colony-forming unit-granulocyte-erythrocyte-macrophage-megakaryocyte [CFU-GEMM]) and erythroid (burst-forming unit-erythroid [BFU-E]) progenitors was assessed in vitro as a potential measure of self-renewal using purified, recombinant (r) human (hu) or murine (mu) mast cell growth factor (MGF), a ligand for the c-kit proto-oncogene receptor. Primary cultures of human umbilical cord blood or adult human bone marrow cells were initiated in methylcellulose with erythropoietin (Epo) alone or in combination with rhu interleukin-3 (IL-3) or MGF. Individual day 14 to 18 CFU-GEMM or BFU-E colonies were removed from primary cultures and reseeded into secondary methylcellulose cultures containing a combination of Epo, MGF, and rhu granulocyte-macrophage colony-stimulating factor (GM-CSF). The data showed a high replating efficiency of cord blood and bone marrow CFU-GEMM in response to Epo + MGF in terms of the percentage of colonies that could be replated and the number of secondary colonies formed per replated primary colony. The average number of hematopoietic colonies and clusters apparent from replated cultures of cord blood or bone marrow CFU-GEMM stimulated by Epo + MGF was greater than with Epo + rhuIL-3 or Epo alone. Replated cord blood CFU-GEMM gave rise to CFU-GEMM, BFU-E, and GM colony-forming units (CFU-GM) in secondary cultures. Replated bone marrow CFU-GEMM gave rise mainly to CFU-GM in secondary cultures. A more limited capacity for replating of cord blood and bone marrow BFU-E was observed. These studies show that CFU-GEMM responding to MGF have an enhanced replating potential, which may be promoted by MGF. These studies also support the concept that MGF acts on more primitive progenitors than IL-3.
利用纯化的重组人（hu）或鼠（mu）肥大细胞生长因子（MGF，一种c-kit原癌基因受体的配体），在体外评估人多能祖细胞（集落形成单位-粒细胞-红细胞-巨噬细胞-巨核细胞[CFU-GEMM]）和红系祖细胞（爆式红系集落形成单位[BFU-E]）的再接种能力，以此作为自我更新的一项潜在指标。人脐带血或成人骨髓细胞的原代培养在甲基纤维素中开始，单独使用促红细胞生成素（Epo），或与重组人白细胞介素-3（IL-3）或MGF联合使用。从原代培养中取出第14至18天的单个CFU-GEMM或BFU-E集落，重新接种到含有Epo、MGF和重组人粒细胞-巨噬细胞集落刺激因子（GM-CSF）组合的二次甲基纤维素培养物中。数据显示，就可再接种的集落百分比以及每个再接种的原代集落形成的二次集落数量而言，脐带血和骨髓CFU-GEMM对Epo + MGF的再接种效率很高。由Epo + MGF刺激的脐带血或骨髓CFU-GEMM再接种培养物中明显可见的造血集落和集簇的平均数量，比Epo + 重组人IL-3或单独使用Epo时更多。再接种的脐带血CFU-GEMM在二次培养中产生CFU-GEMM、BFU-E和GM集落形成单位（CFU-GM）。再接种的骨髓CFU-GEMM在二次培养中主要产生CFU-GM。观察到脐带血和骨髓BFU-E的再接种能力较为有限。这些研究表明，对MGF有反应的CFU-GEMM具有增强的再接种潜力，这可能由MGF促进。这些研究还支持这样的概念，即MGF作用于比IL-3更原始的祖细胞。
