Mar 24, 2016

Cryopreservation of human mucosal leukocytes

BioRxiv : the Preprint Server for Biology
Sean M. HughesFlorian Hladik

Abstract

Background Understanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible. Methods and findings To find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes. Cryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to th...Continue Reading

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Mentioned in this Paper

T-Lymphocyte
Endocervicitis
Isolation Aspects
White Blood Cell Count Procedure
Small Lymphocyte
Cell Count
HIV Infections
Cryopreservation
Stimulation Procedure
Leukocytes

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