Emerson S G, Sieff C A, Wang E A, Wong G G, Clark S C, Nathan D G
J Clin Invest. 1985 Sep;76(3):1286-90. doi: 10.1172/JCI112087.
To facilitate the direct study of progenitor cell biology, we have developed a simple and efficient procedure based upon negative selection by panning to purify large numbers of committed erythroid and myeloid progenitors from human fetal liver. The nonadherent, panned cells constitute a highly enriched population of progenitor cells, containing 30.4 +/- 13.1% erythrocyte burst forming units (BFU-E), 5.5 +/- 1.9% granulocyte-macrophage colony forming units (CFU-GM), and 1.4 +/- 0.7% granulocyte-erythroid-macrophage-megakaryocyte colony forming units (CFU-GEMM) as assayed in methylcellulose cultures. These cells are morphologically immature blasts with prominent Golgi. This preparative method recovers 60-100% of the committed progenitors detectable in unfractionated fetal liver and yields 2-30 X 10(6) progenitors from each fetal liver sample, and thus provides sufficient numbers of enriched progenitors to allow direct biochemical and immunologic manipulation. Using this technique, a purified recombinant protein previously thought to have only granulocyte-macrophage colony stimulating activity (GM-CSA) is shown to have both burst promoting activity and multipotential colony stimulating activity. Progenitor purification by panning thus appears to be a simple, efficient method that should facilitate the direct study of committed hematopoietic progenitors and their differentiation.
为便于直接研究祖细胞生物学,我们开发了一种基于淘选阴性选择的简单高效方法,用于从人胎肝中纯化大量定向红细胞系和髓系祖细胞。未贴壁的淘选细胞构成了祖细胞的高度富集群体,在甲基纤维素培养中检测发现,其中含有30.4±13.1%的红细胞爆式集落形成单位(BFU-E)、5.5±1.9%的粒细胞-巨噬细胞集落形成单位(CFU-GM)以及1.4±0.7%的粒细胞-红细胞-巨噬细胞-巨核细胞集落形成单位(CFU-GEMM)。这些细胞在形态上是未成熟的母细胞,具有明显的高尔基体。这种制备方法能回收未分级胎肝中可检测到的60 - 100%的定向祖细胞,每个胎肝样本可产生2 - 30×10⁶个祖细胞,从而提供了足够数量的富集祖细胞,以便进行直接的生化和免疫操作。利用该技术,一种先前被认为仅具有粒细胞-巨噬细胞集落刺激活性(GM-CSA)的纯化重组蛋白,被证明同时具有爆式促进活性和多能集落刺激活性。因此,通过淘选进行祖细胞纯化似乎是一种简单、高效的方法,应有助于直接研究定向造血祖细胞及其分化。