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Isolation of Leukocytes from the Human Maternal-fetal Interface
08:19

Isolation of Leukocytes from the Human Maternal-fetal Interface

Published on: May 21, 2015

16.7K

A trypsin-based method for isolating leukocytes from human choriodecidua suitable for immunophenotyping and

Karla MacDonald-Ramos1, Marcia Arenas-Hernandez2, Ismael Mancilla-Herrera3

  • 1Departamento de Nutrición y Bioprogramación, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Montes Urales 800, Lomas Virreyes, 11000 Mexico City, Mexico.

Immunobiology
|October 2, 2018
PubMed

Insights

Researchers developed a new method to isolate leukocytes from the maternal-fetal interface. This gentle trypsin digestion technique preserves cell surface markers for flow cytometry and yields high-quality RNA for genetic analysis, aiding labor research.

Area of Science:

  • Reproductive immunology
  • Cellular and molecular biology

Background:

  • Leukocytes at the maternal-fetal interface are crucial in labor inflammation.
  • Current isolation methods (scraping, enzyme digestion) hinder downstream analyses like immunophenotyping and transcriptomics.

Purpose of the Study:

  • To develop an effective leukocyte isolation method from the choriodecidua for advanced analyses.
  • To ensure isolated leukocytes are suitable for flow cytometry and RNA-based techniques.

Main Methods:

  • Gentle trypsin digestion of the choriodecidua.
  • Isolation of leukocyte-enriched cell mixtures.
  • Assessment of lymphocyte and myeloid cell viability.
  • Evaluation of cell surface marker integrity for flow cytometry.
  • RNA extraction for quantitative real-time PCR (qRT-PCR) and microarray analysis.

Main Results:

  • The gentle trypsin digestion method yields a leukocyte-enriched cell population.
  • High viability was observed for lymphocytes, with reduced viability in myeloid cells.
  • Isolated leukocytes maintained cell surface marker integrity, suitable for flow cytometry.
  • High-quality RNA was successfully obtained for qRT-PCR and microarray analyses.

Conclusions:

  • Gentle trypsin digestion is a viable method for isolating leukocytes from the maternal-fetal interface.
  • This method supports downstream immunophenotyping and transcriptomic analyses, crucial for studying labor mechanisms.
  • The technique offers improved cell recovery and suitability for molecular analyses compared to traditional methods.

Keywords:
ChoriodeciduaFlow cytometryIsolationLeukocytesMicroarraysTrypsinqRT-PCR

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