Preparation of Viable Isolated Bovine Parathyroid Cells

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Abstract
A method is described for the preparation of isolated bovine parathyroid cells. Fresh, minced bovine parathyroid tissue is incubated with 2 mg/ml collagenase and 50 .mu.g/ml DNAse for 1 h at 37.degree. C with periodic pipetting. Parathyroid cells are separated from contaminating fat cells and red cells by low speed centrifugation. The resulting cell preparation is indistinguishable from parathyroid cells in intact bovine glands by light and EM. Hormone release is linear for at least 3 h at both high (2.0 mM) and low (0.5 mM) Ca concentrations and is inversely proportional to divalent cation concentration between 0.3 mM and 1.0 mM Ca, as was observed previously both in vivo and in vitro. As in previous studies, hormone release is also stimulated up to 2-fold by 10-6 M (-)isoproterenol, an effect blocked by 10-6 M (-)propranolol, suggesting a .beta.-adrenergic effect. Such a cell preparation should be useful for studying hormone binding and effects on cyclic nucleotides and cellular transport phenomena in both normal and abnormal tissue.

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