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Animal Breeding and Genetics
Asian-Australasian Journal of Animal Sciences 2002;15(2): 274-278.
DOI: https://doi.org/10.5713/ajas.2002.274    Published online January 1, 2002.
Bioluminescent Determination of Lactose Secretion: A Measure of the In Vitro Performance of Mammary Acini from Lactating Rats
B. H. Choi, K. W. Stewart, S. R. Davis, K. H. Myung
Abstract
A culture system for lactating rat mammary acini was evaluated, where the primary indicator of performance was lactose secretion, measured by a sensitive bioluminescence assay. Lactose secretion was reduced by half (p<0.01) over the first 6 h of culture by overnight feed withdrawal (FW) from tissue donors but was sensitive to increased glucose concentration in the culture media (p<0.001) up to 30 mM. Lactose production of cells from fed donors over the first 6 h in culture in 30 mM glucose was 8.9 fmol/cell/h -a rate calculated to be about half that in vivo. No significant difference was shown in lactose secretion by cells from fed or FW rats over 6-24 h. Lactose secretion was 3.6 fmol/cell/h by cells from fed animals in 40 mM glucose concentration media over the 6-24 h culture period. Addition of insulin to the culture media had no effect on rates of lactose secretion while addition of prolactin and hydrocortisone, with or without insulin, significantly (p<0.001) decreased lactose production over both 0-6 h and 6-24 h culture periods. Lactose synthesis in vitro was significantly enhanced by aeration of the media during collagenase digestion of mammary tissue (p<0.05). No improvement in lactose secretion was effected by shaking of cells during culture, Matrigel coating of culture dishes or change in cell density over a range up to 2.5 million cells per ml.
Keywords: Lactose; Lactation; Mammary; Cell Culture; Hormones
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