A lymphocyte colony assay for culture of sheep lymphocytes.
A procedure for culturing sheep peripheral blood lymphocytes in soft agar was standardized. The variables that were evaluated included the type of T lymphocyte mitogens, mitogen doses, cell number, distribution of the cells in the culture vessel and the duration of the culture. Both concanavalin A (CON A) and phytohemagglutinin A (PHA) stimulated the formation of lymphocyte colonies. However, CON A consistently produced more and larger colonies. The doses of CON A and PHA which yielded the maximum number of colonies were 35 and 61 micrograms/ml, respectively. The optimum number of cells per well as 1.36 x 10(6). Adherence of the cells to the floor of the culture vessel was required for the colonies to form. The maximum number of colonies was attained between day 5 and day 6 of culture. The lymphocyte colony assay may be a useful adjunct to other immunologic assays for studying cell-mediated immunity of sheep.
Johnson
R
R
Department of Clinical Sciences, College of Veterinary Medicine, E. Lansing, MI 48824-1316.
Kaneene
J B
JB
Lloyd
J W
JW
D000818:Animals; D002448:Cell Adhesion; D003114:Colony-Forming Units Assay / Q000662:veterinary*; D007958:Leukocyte Count / Q000187:drug effects; D007963:Leukocytes, Mononuclear / Q000502:physiology; D008213:Lymphocyte Activation; D008214:Lymphocytes; D008934:Mitogens / Q000494:pharmacology; D012756:Sheep / Q000097:blood*; D013997:Time Factors
D008934:Mitogens