TY - JOUR
T1 - Culture of Dispersed Anterior Pituitary Cells on Extracellular Matrix
AU - Weiner, Richard I.
AU - Bethea, Cynthia L.
AU - Jaquet, Philippe
AU - Ramsdell, John S.
AU - Gospodarowicz, Denis J.
N1 - Funding Information:
Some of the work in this report was supported in part by Grant CA 23248 from the National Cancer Institute and NASA grants NAS 9-15566 and 2-OR-589-101. The superb secretarial assistance of Mrs. Eileen McConnell is acknowledged.
PY - 1983/1/1
Y1 - 1983/1/1
N2 - This chapter describes methods used in the production of extracellular matrix (ECM), dispersion of anterior pituitary tissue, separation of dispersed cells, and development of a serum-free medium for maintenance of cultured cells. The chapter presents a series of cell dispersion techniques for the anterior pituitary in a variety of species. The chapter focuses on obtaining preparations that were acutely functional rather than on obtaining high yields of cells. For this reason, avoid using proteolytic enzymes, since the use of Viokase destroys cell surface receptors for dopamine in bovine anterior pituitary cells. Dispersed anterior pituitary cells are separated from debris and red blood cells by centrifugation through Ficoll-Hypaque as previously described for the isolation of mouse lymphocytes. The Ficoll-Hypaque is the method of choice based on ease of preparation, cell recovery, and loading capacity.
AB - This chapter describes methods used in the production of extracellular matrix (ECM), dispersion of anterior pituitary tissue, separation of dispersed cells, and development of a serum-free medium for maintenance of cultured cells. The chapter presents a series of cell dispersion techniques for the anterior pituitary in a variety of species. The chapter focuses on obtaining preparations that were acutely functional rather than on obtaining high yields of cells. For this reason, avoid using proteolytic enzymes, since the use of Viokase destroys cell surface receptors for dopamine in bovine anterior pituitary cells. Dispersed anterior pituitary cells are separated from debris and red blood cells by centrifugation through Ficoll-Hypaque as previously described for the isolation of mouse lymphocytes. The Ficoll-Hypaque is the method of choice based on ease of preparation, cell recovery, and loading capacity.
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U2 - 10.1016/S0076-6879(83)03018-9
DO - 10.1016/S0076-6879(83)03018-9
M3 - Article
C2 - 6669038
AN - SCOPUS:0021011367
SN - 0076-6879
VL - 103
SP - 287
EP - 293
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -