Characterization of the cells that migrate from metrial glands of the pregnant mouse uterus during explant culture
Characterization of the cells that migrate from metrial glands of the pregnant mouse uterus during explant culture
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DOI：
10.1016/s0165-0378(96)01008-x
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发表时间：
1997-02-01
影响因子：
3.4
通讯作者：
vandenHeuvel, M
中科院分区：
文献类型：
--
作者：
Croy, BA;McBey, BA;vandenHeuvel, M
粒状子宫腺（GMG）细胞是在妊娠期小鼠子宫中发现的雌激素受体和白细胞介素2 （IL-2）受体阳性的自然杀伤细胞系淋巴细胞。由于技术上的困难，对这些细胞（现在更常被称为子宫NK细胞）的功能研究一直受到限制。细胞分离困难，在培养中不能实现细胞的增殖和分化。1988年，穆赫塔尔和斯图尔特（Cell Tiss）。Res., 253, 413-417)报道了一种从怀孕啮齿动物身上分离的子宫腺的外植体培养方法，该方法产生了几乎纯的uNK细胞群。这一重大技术进步支持了随后大多数啮齿动物uNK细胞的功能和分子研究。然而，通过外植体培养程序分离的细胞的质量尚未确定。细胞化学方法用于鉴定和量化从子宫腺迁移的细胞。妊娠中期，迁移的有核细胞几乎全部（约90%）为NK细胞。在妊娠早期，有相当比例（25%）具有淋巴形态的细胞不能被分配到谱系中。在妊娠第6-16天，通过DNA分离和电泳评估细胞从外植体迁移的活力。在任何时候都有细胞凋亡的证据，即使在培养间隔短至4小时后。平行分析子宫腺的组织学切片，使用末端脱氧转移酶标记检测核碎片，在妊娠第12天之前没有支持显著水平的原位uNK细胞死亡。在外植体培养基中添加雌激素、IL-2、各种细胞外基质、蜕细胞或它们的组合不会导致uNK细胞的体外增殖，并且通常不会延长这些细胞在补充血清或无血清培养基中的短期生存能力。因此，uNK细胞的最佳培养条件仍未确定。(C) 1997爱思唯尔科学爱尔兰有限公司
Granulated metrial gland (GMG) cells are estrogen-receptor and Interleukin 2 (IL-2) receptor positive lymphocytes of the Natural Killer cell lineage found in the murine uterus during pregnancy. Functional studies of these cells, which are now more frequently called uterine NK (uNK) cells, have been limited due to technical difficulties. The cells are difficult to isolate and their proliferation and differentiation have not been achieved in culture. In 1988, Mukhtar and Stewart (Cell Tiss. Res., 253, 413-417) reported a method for explant culture of metrial glands isolated from pregnant rodents that yielded an almost pure population of uNK cells. This major technical advance has supported most of the subsequent functional and molecular studies of rodent uNK cells. However, the quality of the cells isolated by the explant culture procedure has not been established. A cytochemical approach was used to identify and quantify the cells migrating from metrial glands. At midpregnancy, almost all (> 90%) migrating nucleated cells were NK cells. Earlier in gestation, a significant proportion (25%) of cells having lymphoid morphology could not be assigned to the lineage. The viability of cells migrating from explants was assessed by DNA isolation and electrophoresis on days 6-16 of gestation. At all times evidence for apoptosis was found, even after culture intervals as brief as 4 h. Parallel analyses of histological sections of the metrial gland, using terminal deoxytransferase labelling to detect nuclear fragmentation, did not support significant levels of uNK cell death in situ prior to day 12 of gestation. Supplementation of the explant culture medium with estrogen, IL-2, various extracellular matrices, decidual cells or combinations of these did not lead to in vitro proliferation of uNK cells and usually did not extend the short term viability of these cells in serum supplemented or serum free media. Thus, the optimal culture conditions for uNK cells remain undefined. (C) 1997 Elsevier Science Ireland Ltd.