Molecular and functional properties of myeloid stem cell growth factors

1)大鼠脾细胞系RSP-2.P3中性粒细胞生长因子（NeuGF）的纯化及n端氨基酸序列。通过在细胞培养中加入诱导剂优化NeuGF的产生，并通过反相高效液相色谱法纯化NeuGF。通过重组DNA技术，NeuGF的n端序列与前人的推断一致。2)正常人尿液中巨噬细胞生长因子（M<phi>GF）的纯化及原代结构。我们纯化的M<phi>GF的氨基末端序列与其cDNA测序推断的结果一致，但M<phi>GF制剂的氨基酸组成表明，它起源于较大的M<phi>GF mRNA，并通过在190.3位置附近的加工进行了修饰。建立了一个新的细胞系并进行了亚克隆。分离后不久，在这些亚克隆中观察到相对于M<phi>GF的NeuGF产量的差异。然而，在培养一段时间后，亚克隆变得彼此相似。结果表明，尽管单个细胞动态波动，但整体种群的多样性保持平衡。4)纯化的NeuGF和M< φ >GF的生物活性。结果表明，小鼠骨髓干细胞对NeGF和M<phi>GF均有反应，M<phi>GF输注6天后巨噬细胞祖细胞增加，M<phi>GF对腹腔巨噬细胞的激活等。

1) Purification and N-terminal Amino Acid Sequence of Neutrophil Growth Factor (NeuGF) from Rat Spleen Cell Line RSP-2.P3.Production of NeuGF was optimized by adding inducers to the cell culture, and the secreted NeuGF was purified by reverse-phase HPLC. Elucidated N-terminal sequence of the NeuGF was consistent with that deduced by others by recombinant DNA technology.2) Purification and Primary Structure of Macrophage Growth Factor (M<phi>GF) from Normal Human Urine. Amino terminal sequence of the M<phi>GF we purified was consistent with that deduced by others by sequenceing of its cDNA, but amino acid composition of the M<phi>GF preparation suggested that it is originated from the larger-size M<phi>GF mRNA and modified by processing around the position 190.3) Intraclonal Heterogeneity of Mouse Fibrosarcoma Cells That Secrete Both NeuGF and M<phi>GF. A new cell line was established and subcloned. Soon after isolation, dissimilarity was observed among these subclones regarding the production of NeuGF relative to M<phi>GF. However, the subclones became similar to each other when kept in culture for some time. The result suggests that equilibrated diversity is maintained as a whole population although individual cells dynamically fluctuate.4) Biological Activity of the Purified NeuGF and M<phi>GF. Presence of bone marrow stem cells reactive both to NeGF and to M<phi>GF, increase of macrophage progenitor cells after 6-day's infusion of M<phi>GF in mice, activation of peritoneal resident macrophages by M<phi>GF etc. were demonstrated.