FORMULATION OF MEDIUM FOR TICK CELL-CULTURE
FORMULATION OF MEDIUM FOR TICK CELL-CULTURE
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DOI：
10.1007/bf01194061
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发表时间：
1989-08-01
影响因子：
2.2
通讯作者：
KURTTI, TJ
中科院分区：
文献类型：
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作者：
MUNDERLOH, UG;KURTTI, TJ
我们研究了牛胆固醇浓缩物在降低高水平（10-20%）的胎牛血清（FBS）的有效性，这是促进蜱细胞体外生长所必需的。实验采用了从尼氏蜱（Anocentor nitens） （ANE 58）、微嗜牛（Boophilus microplus） （BME 26）和尾鼻蜱（Rhipicephalus appendiculatus） （RAE 25）胚胎分离的蜱细胞系。分别在L-15 （BME 26）或L-15B （ANE 58和RAE 25）培养基中培养，分别添加10%的胰蛋白酶磷酸肉汤（TPB）、5%的ANE 58和BME 26)或3%的牛血清，10-90 .mu。米/毫升胆固醇。浓度为10 μ。G /ml胆固醇对3个品系的生长均有刺激作用，但大于30 .mu。g/ml的胆固醇浓度抑制了ANE 58和RAE 25细胞的生长，而BME 26细胞的增殖则被所有测试的胆固醇浓度增强。3个品系均可在5% FBS条件下连续生长，条件为10.10亩。G /ml胆固醇。在L-15B配方中单独或联合添加L-15的营养物质，在仅添加5% FBS和10 .mu的培养基中测试其支持蜱虫细胞生长的能力。米/毫升胆固醇。单独使用L-15时，rae25细胞不增殖。加入葡萄糖（Glc）、谷氨酸（Glu）或α。-酮戊二酸（. α . k）几乎没有影响，Glc +。α的组合也是如此。K，天冬氨酸（Asp）加脯氨酸（Pro）和谷氨酰胺（Gln），矿物质加维生素（MV）。当Asp， Gln， Pro和。alpha。将K与Glc和/或MV结合，添加到L-15中，对生长有明显的刺激作用，但以Glu同时加入时效果最好。在此培养基中，即L-15B添加5% FBS和10 .mu。/ml胆固醇，BME 26和RAE 25可以连续传代培养。
We examined the effectiveness of bovine cholesterol concentrate in reducing the high level (10-20%) of fetal bovine serum (FBS) necessary to promote tick cell growth in vitro. Tick cell lines isolated from embryos of Anocentor nitens (ANE 58), Boophilus microplus (BME 26), and Rhipicephalus appendiculatus (RAE 25) were used. They were incubated in L-15 (BME 26) or L-15B (ANE 58 and RAE 25) supplemented with 10% tryptose phosphate broth (TPB), 5% (ANE 58 and BME 26) or 3% FBS, 10-90 .mu.m/ml cholesterol. A concentration of 10 .mu.g/ml cholesterol stimulated the growth rate of all three lines but more than 30 .mu.g/ml depressed growth in ANE 58 and RAE 25 cells, while multiplication of BME 26 cells was enhanced by all cholesterol concentrations tested. All three lines could be continuously grown in 5% FBS, provided that 10 .mu.g/ml cholesterol was included. Nutrients added to L-15 in the formulation of L-15B were tested singly or in combination for their ability to support tick cell growth in medium supplemented only with 5% FBS and 10 .mu.m/ml cholesterol. In L-15 alone, RAE 25 cells did not multiply. Adding glucose (Glc), glutamic acid (Glu), or .alpha.-ketoglutaric acid (.alpha.K) had little or no effect, and the same was true for combinations of Glc plus .alpha. K, aspartic acid (Asp) plus proline (Pro) and glutamine (Gln), and minerals plus vitamins (MV). When Asp, Gln, Pro, and .alpha.K were combined with Glc and/or MV and added to L-15, there was appreciable growth stimulation, but best results were obtained when Glu was also included. In this medium, i.e., L-15B with 5% FBS and 10 .mu./ml cholesterol, lines BME 26 and RAE 25 could be continuously subcultured.