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A culture system for the maintenance and proliferation of shark and sting ray immunocytes
Author(s) -
Grogan E. D.,
Lund R.
Publication year - 1990
Publication title -
journal of fish biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.672
H-Index - 115
eISSN - 1095-8649
pISSN - 0022-1112
DOI - 10.1111/j.1095-8649.1990.tb04318.x
Subject(s) - biology , ficoll , incubation , in vitro , andrology , microbiology and biotechnology , immunology , biochemistry , peripheral blood mononuclear cell , medicine
A method designed to support the in vitro maintenance and proliferation of elasmobranch peripheral blood leucocytes (PBLs) has been developed. This method includes protocols for leucocyte isolation, the preparation of media, and culturing conditions. Leucocytes were isolated from whole blood using a Ficoll‐hypaque density gradient followed by nylon wool filtration. Recovered cells were washed and incubated at a density of 0.054 × 10 6 cells per well of a microtitre plate using a novel medium designed for shark cells. Incubation was carried out at 17 and 37° C. These media represent a modification of RPMI 1640 or DMEM to account for isotonic and isoosmotic differences between mammalian and elasmobranch blood via the addition of trimethylamine‐ N ‐oxide, urea and sodium chloride to the RPMI or DMEM.