Characterization of normal peripheral blood lymphocyte colony-forming cells: cell cycle status, surface markers, and cellular growth requirements.
Characterization of normal peripheral blood lymphocyte colony-forming cells: cell cycle status, surface markers, and cellular growth requirements.
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正常外周血淋巴细胞集落形成细胞的特征：细胞周期状态、表面标志物和细胞生长要求。
DOI：
10.1182/blood.v61.3.548.548
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发表时间：
1983
期刊：
影响因子：
20.3
通讯作者：
John Mendelsohn
中科院分区：
文献类型：
--
作者：
R. Taetle;D. To;Alendry P. Caviles;S. Norby;John Mendelsohn
我们进行了一系列的研究，以进一步阐明淋巴细胞集落形成细胞（CFC）的性质，从正常外周血。将单核细胞分离成E-玫瑰花结富集（E+）和E-玫瑰花结耗尽（E-）群体，并在甲基纤维素中与条件培养基和辐照的单核细胞一起培养。获得线性平板关系，其中E+ CFC的平板效率为0.26% +/- .02%（平均值+/- SE），E-CFC的平板效率为0.18% +/- .02%。通过细胞表面标记分析确定，E+集落中的细胞为T淋巴细胞，E-集落中的细胞为B淋巴细胞。用胸腺嘧啶核苷自杀技术，发现约一半的CFC在任何时刻都处于周期中，并且E+ CFC的平板效率和细胞周期状态在液体培养中用PHA预孵育48小时后没有改变.用抗体补体介导的细胞毒性，发现E+ CFC为T101+，OKT 3+，Ia-，而E-CFC为OKT 3-和Ia+.使用通过在单位重力下沉降获得的单核细胞耗尽的群体，淋巴细胞集落生长在单核细胞耗尽的组分中不存在，并且在培养物中40%单核细胞时发生最佳生长。与以前的一些研究相反，我们发现淋巴细胞CFC起源于一个小的，周期性的细胞群体，具有成熟的T或B淋巴细胞标记。然而，进入细胞分裂并不赋予淋巴细胞集落形成能力。单核细胞对E+ CFC的生长至关重要，并且严重耗尽单核细胞的培养物预计不会形成集落。
We performed a series of studies to further clarify the nature of lymphocyte colony-forming cells (CFC) from normal peripheral blood. Mononuclear cells were separated into E-rosette-enriched (E+) and E-rosette-depleted (E-) populations and cultured in methylcellulose with conditioned media and irradiated mononuclear cells. Linear plating relationships were obtained with plating efficiencies of 0.26% +/- .02% (mean +/- SE) for E+ CFC and 0.18% +/- .02% for E- CFC. Cells in E+ colonies were T lymphocytes and in E- colonies were B lymphocytes as determined by cell surface marker analysis. Using the thymidine suicide technique, approximately one-half of CFC were found to be in cycle at any moment, and plating efficiencies and cell cycle status of E+ CFC were not changed by preincubation with PHA in liquid culture for 48 hr. Using antibody complement-mediated cytotoxicity, E+ CFC were found to be T101+, OKT3+, and Ia-, while E- CFC were OKT3- and Ia+. Using monocyte-depleted populations obtained by sedimentation at unit gravity, lymphocyte colony growth was absent in monocyte-depleted fractions, and optimal growth occurred with 40% monocytes in culture. In contrast to some previous studies, we find that lymphocyte CFC originate from a small, cycling population of cells bearing mature T or B lymphocyte markers. Entry into cell division, however, does not confer colony-forming capacity on lymphocytes. Monocytes are critical to growth of E+ CFC, and cultures severely depleted of monocytes would not be expected to form colonies.