Abstract：
(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 CD80+CD163- and CD80-CD163+ cells we could not find differences. Interestingly we found an increased number of double positive cells (CD80+CD163+) 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.
摘要：
(1)研究背景:胎盘免疫细胞在成功形成胎盘和预防妊娠并发症中起着非常重要的作用。巨噬细胞在胎盘的母体和胎儿部分的数量和相关性上占主导地位。胎儿和母体巨噬细胞的极化状态都参与其中的证据，健康和妊娠相关疾病，是有限的。到目前为止，还没有代表性的分离方法可以直接比较母体和胎儿巨噬细胞。（2）材料和方法：从足月胎盘中分离蜕膜巨噬细胞和Hofbauer细胞，机械解剖新鲜组织并用胰蛋白酶和胶原酶A消化。之后,通过Percoll梯度增加细胞富集。CD68表示为泛巨噬细胞标记，进一步研究了表面标志物CD80和CD163。（3）结果：所建立的方法揭示了分离的巨噬细胞的高细胞产率和纯度，并使蜕膜巨噬细胞和Hofbauer细胞之间的比较成为可能。不同巨噬细胞群中单个CD163+细胞的百分比没有观察到显著差异，通过使用FACS和免疫荧光染色。蜕膜巨噬细胞中CD80细胞略有增加。考虑到CD80+CD163-和CD80-CD163+细胞的百分比,我们找不到差异。有趣的是，我们发现与Hofbauer细胞相比，蜕膜巨噬细胞群中的双阳性细胞（CD80CD163）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中的蜕膜巨噬细胞和Hofbauer细胞。它代表了一种有希望的直接细胞比较方法，酶独立，并且不受磁珠的影响，了解胎盘巨噬细胞的功能亚群并确定妊娠相关疾病的治疗靶点。
