Effect of cryopreservation on chemotaxis of lymphocytes

Cryopreservation of cells appears to be a potential method of comparing chemotaxis of lymphocytes collected from different anatomical sites at one time in a single assay. Migration of cryopreserved lymphocytes from swine in the absence (spontaneous migration) or presence of N-formyl-methionyl-leucyl-phenylalanine (fMLP, induced migration) was compared to that of fresh lymphocytes, originating from inguinal (ILN) and mesenteric (MLN) lymph nodes, respectively, using a 48-well microchemotaxis chamber. Cryopreservation did not affect the optimal concentration of fMLP for maximal induced migration and did not impair the chemoattractant activity of fMLP as shown by checkerboard assay. However, cryopreservation reduced the extent of fMLP-induced migration by affecting the spontaneous motility of calls, an effect which was greater for MLN than for ILN cells. This reduction was not related to a loss of cell subset and is in keeping with the view that spontaneous and induced migration involve distinct mechanisms. Thus cryopreservation may be of general use to study the migration of lymphocytes by reducing the differences in spontaneous migration of lymphocytes from different sites.