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Effect of Calcium Concentration on Survival, Proliferation and Activities of Alkaline Phosphatase, 5'-Nucleotidase, γ-Glutamyltransferase and Lactate Dehydrogenase of Adult Rat Hepatocytes Cultured in Serum-free Medium

机译:钙浓度对无血清培养基中成年大鼠肝细胞存活,增殖及碱性磷酸酶,5'-核苷酸酶,γ-谷氨酰转移酶和乳酸脱氢酶活性的影响

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References(22) Cited-By(2) Mature rat hepatocytes were cultured on collagen coated dishes in serum-free a-modified Eagle's minimum essential medium containing 0.1 μM insulin, 0.1 μM dexamethasone, 10 mM pyruvate and Ca2+ at concentrations of 0-2 mM. Survival of nondivided cells was best in medium containing 2 mM Ca2+. Proliferation during 5-day culture was greatest with 0.4 mM Ca2+, but DNA synthesis was scarcely affected by the concentration of Ca2+. Both the activities of alkaline phosphatase, 5 -nucleotidase, γ-glutamyltransferase and lactate dehydrogenase and the number of cell nuclei of cultures in 0.1 mM and 2 mM Ca2+ media were assayed over a 5-day period, and their activities were calculated as enzyme activities per unit number of cell nuclei. Alkaline phosphatase activity increased rapidly during the first day of culture in both media, and its activity in 0.1 mM medium was higher than that in 2 mM medium after culture for 3 days. The activity of 5'-nucleotidase became higher in 0.1 mM medium than in 2 mM medium from day 2 and was maximal on day 3 in both media. ϒ-Glutamyltransferase activity increased and lactate dehydrogenase activity decreased with time in culture, both activities showing no appreciable difference in the two media.
机译:参考文献(22)被引用的By(2)将成熟的大鼠肝细胞在无血清的a-修饰的Eagle最低必需培养基中的胶原蛋白包被培养皿上培养,该培养基含有0.1μM胰岛素,0.1μM地塞米松,10 mM丙酮酸和Ca2 +(浓度为0-2)。毫米在含有2 mM Ca2 +的培养基中,未分裂细胞的存活率最高。用0.4 mM Ca2 +进行5天培养时的增殖最大,但DNA合成几乎不受Ca2 +浓度的影响。在5天的时间内测定了碱性磷酸酶,5-核苷酸酶,γ-谷氨酰转移酶和乳酸脱氢酶的活性以及0.1mM和2mM Ca2 +培养基中培养物的细胞核数目,并将它们的活性计算为酶活性。每单位细胞核数目。在两种培养基中培养的第一天,碱性磷酸酶的活性迅速增加,并且在培养3天后,其在0.1mM培养基中的活性高于在2mM培养基中的活性。从第2天开始,在0.1 mM培养基中5'核苷酸酶的活性高于在2 mM培养基中的活性,并且在两种培养基中的第3天都达到最大值。 β-谷氨酰胺基转移酶的活性随培养时间的延长而增加,乳酸脱氢酶的活性随时间的推移而降低,两种活性在两种培养基中均无明显差异。

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