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

Abstract:

:Leukocytes found at the human maternal-fetal interface participate in the inflammatory process associated with both preterm and term labor; therein, effective methods for their isolation that allow further phenotypic and functional analyses are necessary. Leukocyte isolation is usually carried out through scraping or enzyme digestion of the choriodecidua, however both methods usually limit the use of downstream immunophenotyping or transcriptomic techniques. Here we describe an isolation method based on gentle trypsin digestion that yields a leukocyte-enriched cell mixture with high lymphocyte viability, although less viable myeloid cells. We show that the method does not compromise cell surface markers since isolated leukocytes are suitable for flow cytometry; and that high quality RNA can be obtained from these cells for qRT-PCR and microarray analyses.

journal_name

Immunobiology

journal_title

Immunobiology

authors

MacDonald-Ramos K,Arenas-Hernandez M,Mancilla-Herrera I,Rangel-Escareño C,Vega-Sanchez R

doi

10.1016/j.imbio.2018.09.001

subject

Has Abstract

pub_date

2019-01-01 00:00:00

pages

177-181

issue

1

eissn

0171-2985

issn

1878-3279

pii

S0171-2985(18)30128-1

journal_volume

224

pub_type

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