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{"title":"Isolation of Human Placental Mast Cells","authors":"Soraya Mezouar, Amira Ben Amara, Joana Vitte, Jean-Louis Mege","doi":"10.1002/cpcb.52","DOIUrl":null,"url":null,"abstract":"<p>Mast cells have been identified as resident cells of human placental tissue by immunohistological procedures, suggesting that they may play a role in pregnancy. However, the study of placental mast cells requires their isolation. Here, we describe a procedure to isolate placental mast cells from placenta of healthy women. At-term placentas were recovered, and small pieces were excised. After extensive washing, they were digested using enzyme, and cell preparations were centrifuged on a Percoll density gradient. A double positive selection was then performed using magnetic beads covered with CD117 and IgE antibodies. The purity of isolated mast cells was finally analyzed by flow cytometry, and was nearly 90%, demonstrating that our protocol was convenient to obtain fresh placental mast cells in sufficient quantity for research investigations. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":40051,"journal":{"name":"Current Protocols in Cell Biology","volume":"80 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpcb.52","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpcb.52","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
Mast cells have been identified as resident cells of human placental tissue by immunohistological procedures, suggesting that they may play a role in pregnancy. However, the study of placental mast cells requires their isolation. Here, we describe a procedure to isolate placental mast cells from placenta of healthy women. At-term placentas were recovered, and small pieces were excised. After extensive washing, they were digested using enzyme, and cell preparations were centrifuged on a Percoll density gradient. A double positive selection was then performed using magnetic beads covered with CD117 and IgE antibodies. The purity of isolated mast cells was finally analyzed by flow cytometry, and was nearly 90%, demonstrating that our protocol was convenient to obtain fresh placental mast cells in sufficient quantity for research investigations. © 2018 by John Wiley & Sons, Inc.
人胎盘肥大细胞的分离
肥大细胞已被确定为人类胎盘组织的常驻细胞,通过免疫组织学程序,表明它们可能在妊娠中发挥作用。然而,胎盘肥大细胞的研究需要分离它们。在这里,我们描述了一种从健康妇女胎盘中分离胎盘肥大细胞的方法。恢复足月胎盘,并切除小块。经过大量洗涤后,用酶消化,并用Percoll密度梯度离心细胞制剂。然后用覆盖CD117和IgE抗体的磁珠进行双阳性选择。最后通过流式细胞术对分离的肥大细胞纯度进行了分析,纯度接近90%,表明我们的方法可以方便地获得足够数量的新鲜胎盘肥大细胞进行研究。©2018 by John Wiley &儿子,Inc。
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