Rhyner K, Taetle R
Int J Cell Cloning. 1986 Jan;4(1):27-42. doi: 10.1002/stem.5530040104.
In a previous study, colony-stimulating factor (CSF) activity assayed in colony culture correlated closely with 3HTdR 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 and 3HTdR 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 showed 3HTdR 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-Ia for CFU-gm and liquid culture cells was compared and the majority of both cell populations expressed Ia-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/G1 cells. 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.
在先前的一项研究中，集落培养中检测的集落刺激因子（CSF）活性与去除贴壁细胞的人骨髓细胞对3H-胸苷（3HTdR）的摄取密切相关。为了利用该检测方法筛选CSF培养基和骨髓毒性免疫毒素，将液体培养中的细胞与传统的粒细胞/巨噬细胞（CFU-gm）集落检测进行了比较。液体培养和集落形成检测的CSF剂量反应关系几乎相同。非贴壁骨髓细胞对3HTdR的摄取与CSF剂量相关，并且培养的细胞数量与3HTdR摄取之间存在线性关系。液体培养和CFU-gm的蓖麻毒素细胞毒性曲线在第7天相同，但在第14天培养时显示出一些差异。所有培养物的结果表明，3HTdR摄取与CFU-gm集落生长最密切相关，而不是集簇生长。液体培养中的髓细胞分化与集落培养相似，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，可以定量各种髓细胞的产生。比较了抗Ia对CFU-gm和液体培养细胞的细胞毒性，两种细胞群体中的大多数都表达Ia样抗原。同时对表面抗原和DNA含量进行染色以表征增殖的骨髓细胞，绝大多数细胞表达髓系标志物。转铁蛋白受体由处于S/G2/M期的细胞显示，并在CSF刺激后出现在G0/G1期细胞上。我们得出结论，液体培养可用于筛选人CSF的条件培养基，并筛选对正常髓系前体细胞的细胞毒性。液体培养中CSF反应性细胞的行为似乎与CFU-gm集落形成细胞的行为最密切相关，并且对CSF刺激细胞的表征允许对髓细胞产生进行定量和定性估计。
