CSF-responsive bone marrow cells in liquid culture: characterization and screening applications.
CSF-responsive bone marrow cells in liquid culture: characterization and screening applications.
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液体培养中的 CSF 反应性骨髓细胞：表征和筛选应用。
DOI：
10.1002/stem.5530040104
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Taetle,R
中科院分区：
文献类型：
--
作者：
Rhyner,K;Taetle,R
在先前的研究中，在集落培养物中测定的集落刺激因子（CSF）活性与去除粘附细胞的人骨髓细胞的3 HTdR摄取密切相关。为了使用该试验筛选CSF培养基和骨髓毒性免疫毒素，将液体培养物中生长的细胞与常规粒细胞/巨噬细胞（CFU-gm）菌落试验进行比较。液体和集落形成试验的CSF剂量反应关系几乎相同。非粘附骨髓细胞的3 HTdR摄取与CSF剂量相关，培养细胞数量与3 HTdR摄取之间存在线性关系。液体培养物和CFU-gm的蓖麻毒素细胞毒性曲线在第7天相同，但与第14天培养物显示出一些差异。所有培养物的结果均显示3 HTdR摄取与CFU-gm菌落而非簇生长最密切相关。液体培养中的髓样细胞分化类似于集落培养，产生粒细胞、巨噬细胞和嗜酸性粒细胞的混合物。通过结合细胞计数和分类计数，可以定量各种骨髓细胞的产生。比较了抗la对CFU-gm和液体培养细胞的细胞毒性，两种细胞群的大多数表达la样抗原。表面抗原和DNA含量的同时染色用于表征增殖的骨髓细胞，并且绝大多数细胞表达髓样标志物。转铁蛋白受体在S/G2/M期细胞中显示，在G 0/G1期细胞经CSF刺激后出现。我们的结论是液体培养可用于筛选条件培养基的人CSF和筛选细胞毒性正常骨髓前体细胞。液体培养中CSF反应细胞的行为似乎与CFU-gm集落形成细胞的行为最密切相关，并且CSF刺激细胞的表征允许定量和定性估计髓样细胞产生。
In a previous study, colony‐stimulating factor (CSF) activity assayed in colony culture correlated closely with3HTdR uptake by human marrow cells depleted of adherent cells. To use this assay for screening media for CSF and immunotoxins for marrow toxicity, cells growing in liquid culture were compared to conventional granulocyte/macrophage (CFU‐gm) colony assays. CSF dose‐response relationships for liquid and colony‐forming assays were nearly identical.3HTdR uptake by nonadherent marrow cells was CSF dose‐related, and there was a linear relationship between number of cells cultured and3HTdR uptake. Ricin cytotoxicity curves for liquid cultures and CFU‐gm were identical on day 7 but showed some disparity with day 14 cultures. Results with all cultures showed3HTdR uptake to be most closely correlated with CFU‐gm colony, rather than cluster, growth. Myeloid cell differentiation in liquid culture was similar to colony cultures, producing mixtures of granulocytes, macrophages and eosinophils. By combining cell and differential counts, production of various myeloid cells could be quantitated. Cytotoxicity of anti‐la for CFU‐gm and liquid culture cells was compared and the majority of both cell populations expressed la‐like antigens. Simultaneous staining for surface antigens and DNA content was used to characterize proliferating marrow cells, and the vast majority of cells expressed myeloid markers. Transferrin receptors were displayed by cells in S/G2/M and appeared after CSF stimulation on G0/G1cells. We conclude liquid cultures can be used to screen conditioned media for human CSF and to screen for cytotoxicity to normal myeloid precursor cells. Behavior of CSF‐responsive cells in liquid culture appears most closely related to that of CFU‐gm colony‐forming cells, and characterization of CSF‐stimulated cells allows quantitative as well as qualitative estimates of myeloid cell production.
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