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Immunobiology. 2019 Jan;224(1):177-181. doi: 10.1016/j.imbio.2018.09.001. Epub 2018 Sep 22.

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

Author information

1
Departamento de Nutrición y Bioprogramación, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Montes Urales 800, Lomas Virreyes, 11000 Mexico City, Mexico. Electronic address: kmmacdonald@yahoo.com.
2
Departamento de Nutrición y Bioprogramación, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Montes Urales 800, Lomas Virreyes, 11000 Mexico City, Mexico; Departamento de Biomedicina Molecular Centro de Investigación y de Estudios Avanzados, Instituto Politécnico Nacional Av Instituto Politécnico Nacional 2508, La Laguna Ticoman, 07360 Mexico City, Mexico. Electronic address: march.arenas@gmail.com.
3
Departamento de Infectología e Inmunología, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Montes Urales 800, Lomas Virreyes, 11000 Mexico City, Mexico. Electronic address: mahi_25803@yahoo.com.mx.
4
Departamento de Genómica Computacional, Instituto Nacional de Medicina Genómica, Periférico Sur 4809, Arenal Tepepan, 14610 Mexico City, Mexico. Electronic address: crangel@inmegen.gob.mx.
5
Departamento de Nutrición y Bioprogramación, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Montes Urales 800, Lomas Virreyes, 11000 Mexico City, Mexico. Electronic address: vegarodrig@gmail.com.

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.

KEYWORDS:

Choriodecidua; Flow cytometry; Isolation; Leukocytes; Microarrays; Trypsin; qRT-PCR

PMID:
30269980
DOI:
10.1016/j.imbio.2018.09.001
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