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Molecular and functional properties of myeloid stem cell growth factors

骨髓干细胞生长因子的分子和功能特性

基本信息

批准号：
60480465
负责人：
SHIKITA Mikio
金额：
$ 3.01万
依托单位：
National Institute of Radiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480465/
关键词：
Cell Growth Factor Colony-Stimulating Factor Macrophage Neutrophil Cell Culture Cell Line Fibrosarcoma Leukemia Radiation 細胞培養細胞株化

项目摘要

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.

1)大鼠脾细胞系RSP-2.P3中神经生长因子（NeuGF）的纯化及N-末端氨基酸序列分析。通过添加诱导剂优化细胞培养物中NeuGF的产生，并通过反相HPLC纯化分泌的NeuGF。N-端序列的测定结果与其他人用重组DNA技术推导的结果一致。2）正常人尿中巨噬细胞生长因子（M GF）的纯化及一级结构<phi>。我们纯化的M GF的氨基端序列<phi>与其他人通过其cDNA测序推断的序列一致，但M GF制备物的氨基酸组成<phi>表明它来自较大的M <phi>GF mRNA，并在190位附近经过加工修饰。3）分泌NeuGF和M GF的小鼠纤维肉瘤细胞的克隆内异源性<phi>。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NeuGF相对于MGF的生产<phi>的差异。然而，当在培养物中保持一段时间时，亚克隆变得彼此相似。结果表明，尽管单个细胞动态波动，但作为整个群体保持平衡的多样性。4）纯化的NeuGF和MGF的生物活性<phi>。结果表明，小鼠骨髓间充质干细胞对NeGF和MGF均具有反应<phi>性，MGF输注6天后巨噬细胞祖细胞增多<phi>，MGF可激活腹腔巨噬细胞<phi>等。