原代CFU-C菌落细胞的再培养。

G E Hübner, F Ali-Osman, M Kastner, C Papadimitriou, H R Maurer
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摘要

本研究旨在探讨CFU-C衍生菌落的细胞能否形成继代菌落。用含有粒细胞/巨噬细胞集落刺激因子(GM-CSF)的小鼠肺条件培养基(MLCM)刺激玻璃毛细血管中体积在25微升至75微升之间的琼脂培养基培养骨髓。将大于或等于20个菌落的琼脂凝胶吹入相同的培养基中,在旋转混合上完全分散到单细胞悬浮液中,并用于建立二次琼脂培养。在这些继发培养中,出现了相当数量的继发粒细胞、混合粒细胞/巨噬细胞和巨噬细胞菌落以及许多簇。相反,当单菌落再培养时,只形成少量的次生细胞聚集体。当含有大于或等于20个细胞聚集体的原代培养物在3天和4天的间歇时间内进行连续再培养时,通过7天的集落计数监测,发现在第三代培养物中集落形成细胞增加了2-7倍。在不同含csf培养基中,用牛肺条件培养基(BLCM)进行原代培养,用l细胞条件培养基(LCCM)进行继代培养,继代菌落比原代培养增加4倍以上。原代毛细血管培养未发现CFU-S。在培养皿中培养骨髓,用MLCM刺激,获得了原代和继代菌落的生长。结果表明CFU-C在体外具有一定的自我更新能力。
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Reculturing of cells from primary CFU-C colonies.

This study was aimed at investigating whether cells of CFU-C derived colonies could form secondary colonies. Bone marrow cultures of volumes of agar medium between 25 microliter and 75 microliter contained in glass capillaries were stimulated with mouse lung-conditioned medium (MLCM) containing granulocyte/macrophage colony-stimulating factor (GM-CSF). Agar gels with colonies of up to greater than or equal to 20 were blown out into identical culture medium, completely dispersed on a whirl-mix to single cell suspensions, and used for establishing secondary agar cultures. In these secondary cultures considerable numbers of secondary granulocytic, mixed granulocytic/macrophage and macrophage colonies as well as numerous clusters arose. In contrast, when single colonies were recultured, only few secondary cell aggregates were formed. When primary cultures containing up to greater than or equal to 20 cell aggregates were used for serial reculture at intermittent intervals of 3 and 4 days, a 2-7-fold increase of colony-forming cells was found in tertiary cultures as was monitored by 7 day colony counts. And by use of different kinds of CSF-containing media, an over 4-fold increase of secondary over primary colonies was obtained with bovine lung-conditioned medium (BLCM) in primary and L-cell-conditioned medium (LCCM) in secondary 7 day cultures. Primary capillary cultures were found to be devoid of CFU-S. Also, setting up bone marrow cultures in petri dishes and stimulating with MLCM, growth of primary as well as secondary colonies was obtained. The results indicate some self-renewal potential of CFU-C in vitro.

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