Optimal conditions for in vitro blastogenesis of feline peripheral blood lymphocytes

doi:10.1016/0165-2427(82)90014-9

Veterinary Immunology and Immunopathology

Volume 3, Issue 5, September 1982, Pages 485-498

https://doi.org/10.1016/0165-2427(82)90014-9 Get rights and content

Abstract

A microculture technique was developed for the in vitro blastogenesis of feline lymphocytes. Blastogenesis of ficoll-diatriazoate gradient separated mononuclear cell, washed blood and whole blood were compared. In general the whole blood cultures yielded higher stimulation indices (SI) than the washed blood or separated mononuclear cell cultures.

The effect of several variables on the stimulation of lymphocyte cultures was examined. A cell concentration of 3 × 10⁵ cells/well and a 1:20 dilution of washed and unwashed whole blood gave optimal stimulation with concanavalin A (Con A). Phytohaemagglutinin-P (PHA-P) did not give significant levels of stimulation. Inactivated fetal calf serum (FCS) at levels of 2.5% (for washed blood) and 5% (for separated mononuclear cell and whole blood) gave highest SI. Supplementation with FCS was preferable to autologous, homologous or horse sera for all cultures. Optimal SI was obtained in all cultures incubated for 3 days and labelled with 1 μCi tritiated thymidine (³H-TdR) for the last 16 hours. The highest SI were in the range of 70 to 105 (18,764 to 42,681 counts per minute (CPM) for separated mononuclear cell culture, 100 to 165 (28,403 to 45,334 CPM) for washed blood culture and 105 to 186 (41,076 to 69,999 CPM) for whole blood culture.

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Optimal conditions for in vitro blastogenesis of feline peripheral blood lymphocytes

Abstract

Vet. Immunol. Immunopathol.

Res. Vet. Sci.

Vet. Immunol. Immunopathol.

Lancet

Res. Vet. Sci.

Res. Vet. Sci.

J. Immunol. Methods

J. Immunol. Methods

The effects of autologous plasma on human lymphocyte transformation

P.S.E.B.M.

The effect of antibody on antigen-induced lymphocyte transformation

J. Immunol.

Phytomitogen- and antigen-induced blast transformation of feline lymphocytes

Am. J. Vet. Res.

Dissociation and inhibition of the stimulatory effect of phytohaemagglutinin on protein and DNA synthesis in human lymphocyte cultures

Nature

Role of cell-cell interactions in mitogenic stimulation of lymphocytes 1. Dissociation of concanavalin A-inducible lymphocyte activation and agglutination

Journal of Immunogenetics

The role of two distinct accessory cells in T lymphocyte activation by concanavalin A. Kitasato

Arch. of Exp. Med.

Simplified whole blood method for evaluating in vitro lymphocyte reactivity of laboratory animals

Clin. exp. Immunol.

Allograft immunity in vitro 1. Cultivation conditions and mixed lymphocyte interaction of mouse peripheral lymphocytes

Clin. exp. Immunol.

Macrophage-lymphocyte interaction in the antigen-induced blastogenic response of human peripheral blood leukocytes

J. Immunol.