Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital, Ludwig-Maximillians-Universität München, 81377 Munich, Germany.
Walter-Brendel-Centre of Experimental Medicine, University Hospital, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Int J Mol Sci. 2022 May 30;23(11):6113. doi: 10.3390/ijms23116113.
(1) Background: Placental immune cells are playing a very important role in a successful placentation and the prevention of pregnancy complications. Macrophages dominate in number and relevance in the maternal and the fetal part of the placenta. The evidence on the polarization state of fetal and maternal macrophages involved in both, healthy and pregnancy-associated diseases, is limited. There is no representative isolation method for the direct comparison of maternal and fetal macrophages so far. (2) Material and Methods: For the isolation of decidual macrophages and Hofbauer cells from term placenta, fresh tissue was mechanically dissected and digested with trypsin and collagenase A. Afterwards cell enrichment was increased by a Percoll gradient. CD68 is represented as pan-macrophage marker, the surface markers CD80 and CD163 were further investigated. (3) Results: The established method revealed a high cell yield and purity of the isolated macrophages and enabled the comparison between decidual macrophages and Hofbauer cells. No significant difference was observed in the percentage of single CD163 cells in the distinct macrophage populations, by using FACS and immunofluorescence staining. A slight increase of CD80 cells could be found in the decidual macrophages. Considering the percentage of CD80CD163 and CD80CD163 cells we could not find differences. Interestingly we found an increased number of double positive cells (CD80CD163) in the decidual macrophage population in comparison to Hofbauer cells. (4) Conclusion: In this study we demonstrate that our established isolation method enables the investigation of decidual macrophages and Hofbauer cells in the placenta. It represents a promising method for direct cell comparison, enzyme independently, and unaffected by magnetic beads, to understand the functional subsets of placental macrophages and to identify therapeutic targets of pregnancy associated diseases.
（1）背景：胎盘免疫细胞在成功的胎盘形成和预防妊娠并发症中起着非常重要的作用。巨噬细胞在数量和相关性上在胎盘的母体和胎儿部分占主导地位。关于参与健康和与妊娠相关疾病的胎儿和母体巨噬细胞极化状态的证据有限。目前还没有用于直接比较母体和胎儿巨噬细胞的代表性分离方法。（2）材料和方法：为了从足月胎盘中分离蜕膜巨噬细胞和 Hofbauer 细胞，新鲜组织用胰蛋白酶和胶原酶 A 进行机械解剖和消化。然后通过 Percoll 梯度增加细胞富集。CD68 作为巨噬细胞的泛标志物，进一步研究了表面标志物 CD80 和 CD163。（3）结果：所建立的方法显示出分离的巨噬细胞的高细胞产量和纯度，并能够比较蜕膜巨噬细胞和 Hofbauer 细胞。通过流式细胞术和免疫荧光染色，在不同巨噬细胞群体中，单个 CD163 细胞的百分比没有观察到显著差异。在蜕膜巨噬细胞中可以发现 CD80 细胞略有增加。考虑到 CD80CD163 和 CD80CD163 细胞的百分比，我们没有发现差异。有趣的是，我们发现与 Hofbauer 细胞相比，蜕膜巨噬细胞群体中双阳性细胞（CD80CD163）的数量增加。（4）结论：在这项研究中，我们证明了我们建立的分离方法能够研究胎盘的蜕膜巨噬细胞和 Hofbauer 细胞。它代表了一种有前途的方法，可以直接进行细胞比较，不受酶的影响，也不受磁珠的影响，以了解胎盘巨噬细胞的功能亚群，并确定与妊娠相关疾病相关的治疗靶点。
