Croy B A, McBey B A, Villeneuve L A, Kusakabe K, Kiso Y, van den Heuvel M
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Canada.
J Reprod Immunol. 1997 Feb;32(3):241-63. doi: 10.1016/s0165-0378(96)01008-x.
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.
颗粒状蜕膜腺（GMG）细胞是在妊娠期间小鼠子宫中发现的自然杀伤细胞谱系中的雌激素受体和白细胞介素2（IL-2）受体阳性淋巴细胞。由于技术困难，对这些细胞（现在更常被称为子宫自然杀伤（uNK）细胞）的功能研究受到限制。这些细胞难以分离，并且在培养中尚未实现其增殖和分化。1988年，Mukhtar和Stewart（《细胞与组织研究》，253，413 - 417）报道了一种从怀孕啮齿动物分离的蜕膜腺外植体培养方法，该方法产生了几乎纯的uNK细胞群体。这一重大技术进展支持了随后大多数关于啮齿动物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质量尚未确定。采用细胞化学方法来鉴定和量化从蜕膜腺迁移的细胞。在妊娠中期，几乎所有（> 90%）迁移的有核细胞都是NK细胞。在妊娠早期，相当比例（25%）具有淋巴细胞形态的细胞无法归为该谱系。在妊娠第6 - 16天，通过DNA分离和电泳评估从外植体迁移的细胞的活力。在所有时间都发现了凋亡证据，即使培养间隔短至4小时。使用末端脱氧转移酶标记检测核碎片对蜕膜腺组织切片进行的平行分析不支持在妊娠第12天之前原位uNK细胞有显著水平的死亡。用雌激素、IL - 2、各种细胞外基质、蜕膜细胞或这些物质的组合补充外植体培养基不会导致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活力。因此，uNK细胞最佳培养条件仍未明确。
