Goldberg J, McGuire L A, Williams W J
Blood. 1981 Mar;57(3):497-504.
Human blood mononuclear cells were prepared from peripheral blood or single donor plateletpheresis residues by depletion of adherent cells and T lymphocytes. In double-layer soft agar culture, 5 x 10(5) such cells yielded from 33 to 165 colonies. For liquid culture, these cells were suspended in McCoy's 5A medium with 15% fetal bovine serum and 0%, 20%, or 40% conditioned medium (CM) and incubated for up to 14 days at 37 degrees C in a humidified 5% CO2-95% air atmosphere. The number of cells in cultures with CM decreased about 0%-10%, while cell counts from cultures without CM decreased about 45%-65%. In cultures with CM, 5%-20% of the cells were classified as blasts after 3-5 days. After 7-11 days, blasts and promyelocytes comprised up to 53% of all cells. After 9-11 days, cells with specific granules and maturing nuclei comprised up to 56% of all cells. At 11 days, up to 66% of the cells contained peroxidase-positive granules. Cultures without CM contained no more than 5% blasts and promyelocytes and less than 5% maturing granulocytic cells. 3H-thymidine and Na235SO4 incorporation reached a peak at 3-5 days and at 5-11 days, respectively, in cells from cultures containing CM.
人血单核细胞是通过去除贴壁细胞和T淋巴细胞，从外周血或单供体血小板分离残余物中制备的。在双层软琼脂培养中，5×10⁵个这样的细胞可产生33至165个集落。对于液体培养，将这些细胞悬浮于含有15%胎牛血清和0%、20%或40%条件培养基（CM）的 McCoy's 5A培养基中，在37℃、5%二氧化碳 - 95%空气的湿润环境中孵育长达14天。含有CM的培养物中的细胞数量减少约0% - 10%，而不含CM的培养物中的细胞数量减少约45% - 65%。在含有CM的培养物中，3 - 5天后5% - 20%的细胞被分类为原始细胞。7 - 11天后，原始细胞和早幼粒细胞占所有细胞的比例高达53%。9 - 11天后，具有特异性颗粒和成熟细胞核的细胞占所有细胞的比例高达56%。在11天时，高达66%的细胞含有过氧化物酶阳性颗粒。不含CM的培养物中原始细胞和早幼粒细胞不超过5%，成熟粒细胞细胞少于5%。在含有CM的培养物的细胞中，³H - 胸腺嘧啶核苷和Na₂³⁵SO₄掺入分别在3 - 5天和5 - 11天达到峰值。
