Lipid metabolism in biotin-responsive multiple carboxylase deficiency.
Lipid metabolism in biotin-responsive multiple carboxylase deficiency.
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生物素响应性多重羧化酶缺乏症中的脂质代谢。
DOI：
10.1007/bf01805432
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发表时间：
1985
影响因子：
4.2
通讯作者：
Packman,S
中科院分区：
文献类型：
--
作者：
Gonzalez-Rios,MC;Whitney,SC;Williams,ML;Elias,PM;Packman,S
材料和方法成纤维细胞如所述增殖（Packman等，1982 a），其中突变细胞来源于先前表征的全羧化酶合成酶缺陷型患者（Packman等，1981，1982 b），对照细胞（GM 0969和GM 0041）获自Hmnaan Genetic Mutant Cell Repository（Camden）。对于掺入研究，将细胞接种（37 ℃，8%CO2）在DME-H21培养基中，加入L-谷氨酰胺、青霉素-链霉素、透析的胎牛血清（至15%终浓度）、丙酮酸钠（至1 mM终浓度）和补充的生物素（终浓度100 gEl-1）或抗生物素蛋白（终浓度40 U1 -1）。经透析的胎牛血清含有2 -4gEl-1生物素，在含有未经透析的胎牛血清的完全培养基中，生物素浓度为0.3- 0.6gEl-1（Packman等人，1982年a）。一旦达到30-40 °汇合，继续如上生长，除了用脂蛋白和血清蛋白缺乏的胎牛血清替代。在75%汇合时，将纯化的猪胰岛素（Eli Lilly and Co.，1 mUm 1 -1终浓度）加入培养基中，在胰岛素存在下孵育第三天期间，在24小时内测量放射性同位素掺入（Shafrir和Bierman，1981）。
MATERIALS AND METHODS Fibroblasts were propagated as described (Packman et al., 1982a), with mutant cells derived from holocarboxylase synthetase-deficient patients previously characterized (Packman et al, 1981, 1982b) and control cells (GM0969 and GM0041) obtained from the Hmnaan Genetic Mutant Cell Repository (Camden). For incorporation studies, cells were plated (37 C, 8% CO2) in DME-H21 medium, with added L-glutamine, penicillin-streptomycin, dialyzed fetal calf serum (to 15% final concentration), Na pyruvate (to 1 mM final concentration), and either supplemental biotin (final concentration 100gEl-1) or avidin (40U1-1 final concentration). Dialyzed fetal calf serum contains2-4gEl-1 biotin, giving a biotin concentration of 0.3-0.6 gEl-1 in the complete medium containing unsupptemented dialyzed fetal calf serum (Packman et al., 1982a). Once 30-40~ o confluence was reached, growth was continued as above, except for the substitution of lipoprotein-and serum protein-poor fetal calf serum. At 75% confluence, purified pork insulin (Eli Lilly and Co., 1 mUm1-1 final concentration) was added to the medium, and radioisotope incorporation was measured over 24 h during the third day of incubation in the presence of insulin (Shafrir and Bierman, 1981).