Isolation of Human Placental Mast Cells.

Curr Protoc Cell Biol

Aix-Marseille University, INSERM-1095, CNRS-7278, IRD-198, URMITE, Marseille, France.

Published: September 2018

AI Article Synopsis

  • Mast cells are found in human placental tissue and may influence pregnancy, but their study requires isolation.
  • The process described involves retrieving placentas from healthy women and chopping them into pieces, followed by enzyme digestion and centrifugation using a Percoll density gradient.
  • A double selection method with magnetic beads targeting specific antibodies resulted in a 90% purity of isolated mast cells, providing a useful protocol for further research.

Article 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.

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Source
http://dx.doi.org/10.1002/cpcb.52DOI Listing

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