CN110713972A - 一种用于st细胞培养及相应病毒生产的低血清培养基 - Google Patents

一种用于st细胞培养及相应病毒生产的低血清培养基 Download PDF

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CN110713972A
CN110713972A CN201910900412.3A CN201910900412A CN110713972A CN 110713972 A CN110713972 A CN 110713972A CN 201910900412 A CN201910900412 A CN 201910900412A CN 110713972 A CN110713972 A CN 110713972A
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房圆瑗
齐智
刘海英
孟丹丹
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Shandong Oracle Biotechnology Co Ltd
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Abstract

本发明公开了一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：氨基酸、维生素、无机盐类、辅助成分和蛋白质；所述氨基酸包括甘氨酸、盐酸精氨、谷氨酰胺、盐酸赖氨酸、酪氨酸二钠盐二水合物；所述蛋白质包括重组胰岛素、4‑羟乙基哌嗪乙磺酸（HEPES）；本发明制备的用于ST细胞培养及相应病毒生产的低血清培养基中，使用甲骨文生物ST低血清培养基IV系列添加3%猪瘟专用新生牛血清情况下能够替代传统细胞培养基对ST细胞进行常规细胞培养，且细胞培养状态与数量均与传统培养方法无显著差异，保持细胞良好的生长状态；同时利用该培养体系可以较好地增殖猪瘟病毒，病毒滴度不小于50万倍RID。

Description

一种用于ST细胞培养及相应病毒生产的低血清培养基
技术领域
本发明涉及生物制药疫苗生产领域，具体是一种用于ST细胞培养及相应病毒生产的低血清培养基。
背景技术
传统ST细胞培养的培养方式为MEM培养基添加10％小牛血清培养细胞，血清用量大，成本高。
由于ST细胞在培养过程中对血清比较依赖，降低血清培养会出现细胞生长慢、细胞死亡等情况；市面上ST低血清培养基在普通基础培养基基础上添加成分种类多，成分复杂。进行猪瘟病毒生产时，接毒后细胞贴壁不牢，难以稳定生产，且产毒效果差，未达到半成品质量要求；基于此，我们提出一种用于ST细胞培养及相应病毒生产的低血清培养基。
发明内容
本发明的目的在于提供一种用于ST细胞培养及相应病毒生产的低血清培养基，以解决上述背景技术中提出的问题。
为实现上述目的，本发明提供如下技术方案：
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：氨基酸、维生素、无机盐类、辅助成分和蛋白质；所述氨基酸包括甘氨酸5-10mg/L、丙氨酸5-10mg/L、盐酸精氨110-150mg/L、一水天门冬酰10-15mg/L、天门冬氨10-15mg/L、二盐酸胱氨酸20-40mg/L、谷氨酸10-15mg/L、谷氨酰胺200-350mg/L、一水盐酸组氨酸25-50mg/L、异亮氨酸25-60mg/L、亮氨酸25-60mg/L、盐酸赖氨酸50-80mg/L、甲硫氨酸10-20mg/L、苯丙氨酸20-40mg/L、脯氨酸10-15mg/L、丝氨酸10-15mg/L、苏氨酸25-50mg/L、色氨酸5-15mg/L、酪氨酸二钠盐二水合物30-60mg/L、缬氨酸25-50mg/L、羟脯氨酸5-10mg/L；所述蛋白质包括人转铁蛋白5-10mg/L、重组胰岛素5-10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)1500-3400mg/L。
作为本发明进一步的方案：所述维生素包括生物素0.05-0.2mg/L、氯化胆碱
2-5mg/L、D-泛酸钙2-5mg/L、叶酸2-5mg/L、烟酰胺2-5mg/L、吡哆素盐酸盐0.1-0.3mg/L、盐酸吡哆醛0.1-0.3mg/L、核黄素0.1-0.3mg/L、盐酸硫胺素1-5mg/L、B12维生素0.1-0.3mg/L、i-肌醇5-10mg/L、维生素A醋酸酯0.01-0.05mg/L、维生素D
0.01-0.1mg/L、维生素K 0.005-0.01mg/L。
作为本发明再进一步的方案：所述无机盐类包括无水氯化钙150-200mg/L、无水硫酸镁50-120mg/L、氯化钾300-400mg/L、碳酸氢钠2500-3700mg/L、氯化钠5000-6000mg/L、一水磷酸二氢钠120-150mg/L、五水硫酸铜0.0005-0.001mg/L、七水合硫酸亚铁0.1-0.5mg/L、无水氯化镁10-30mg/L。
作为本发明再进一步的方案：所述辅助成分包括D-葡萄糖2000-5000mg/L、乙醇胺1-5mg/L、次黄嘌呤钠0.1-0.5mg/L、亚油酸0.1-0.5mg/L、酚红10-15mg/L、腐胺二盐酸2-6mg/L、丙酮酸钠90-120mg/L、还原型谷胱甘肽0.5-2.0mg/L、半乳糖100-300mg/L、腺嘌呤5-15mg/L、血清素5-15mg/L、水溶性胆固醇1-5mg/L、盐酸鸟嘌呤10-20mg/L、胸腺嘧啶0.1-0.5mg/L、尿嘧啶0.1-0.5mg/L、胸腺嘧啶核苷0.1-0.5mg/L。
作为本发明再进一步的方案：包括以下组成成分：甘氨酸7.5mg/L、丙氨酸8.9mg/L、盐酸精氨酸126mg/L、一水天门冬酰胺13.2mg/L、天门冬氨酸13.3mg/L、二盐酸胱氨酸31.28mg/L、谷氨酸14.7mg/L、谷氨酰胺292mg/L、一水盐酸组氨酸42mg/L、异亮氨酸52mg/L、亮氨酸52mg/L、盐酸赖氨酸72.5mg/L、甲硫氨酸15mg/L、苯丙氨酸32mg/L、脯氨酸11.5mg/L、丝氨酸10.5mg/L、苏氨酸48mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物51.9mg/L、缬氨酸46mg/L、羟脯氨酸6.8mg/L、生物素0.15mg/L、氯化胆碱3.5mg/L、D-泛酸钙3.5mg/L、叶酸3.5mg/L、烟酰胺3.5mg/L、吡哆素盐酸盐0.2mg/L、盐酸吡哆醛0.1mg/L、核黄素0.25mg/L、盐酸硫胺素3.5mg/L、B12维生素0.2mg/L、i-肌醇6.3mg/L、维生素A醋酸酯0.05mg/L、维生素D0.07mg/L、维生素K 0.008mg/L、无水氯化钙180mg/L、无水硫酸镁92mg/L、氯化钾380mg/L、碳酸氢钠2700mg/L、氯化钠5600mg/L、一水磷酸二氢钠130mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.3mg/L、无水氯化镁25mg/L、D-葡萄糖4000mg/L、乙醇胺2.5mg/L、次黄嘌呤钠0.4mg/L、亚油酸0.2mg/L、酚红12mg/L、腐胺二盐酸5.5mg/L、丙酮酸钠105mg/L、还原型谷胱甘肽1.7mg/L、半乳糖200mg/L、腺嘌呤12.5mg/L、血清素5.5mg/L、水溶性胆固醇2.8mg/L、盐酸鸟嘌呤14.5mg/L、胸腺嘧啶0.2mg/L、尿嘧啶0.2mg/L、胸腺嘧啶核苷0.38mg/L、人转铁蛋白6.5mg/L、重组胰岛素10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)2800mg/L。
作为本发明再进一步的方案：所述人转铁蛋白为饱和铁人转铁蛋白。
作为本发明再进一步的方案：所述重组胰岛素为全链重组胰岛素。
与现有技术相比，本发明的有益效果是：
本发明制备的用于ST细胞培养及相应病毒生产的低血清培养基中，使用甲骨文生物ST低血清培养基IV系列添加3％猪瘟专用新生牛血清情况下能够替代传统细胞培养基对ST细胞进行常规细胞培养，且细胞培养状态与数量均与传统培养方法无显著差异，保持细胞良好的生长状态；同时利用该培养体系可以较好地增殖猪瘟病毒，病毒滴度不小于50万倍RID。
附图说明
图1为MEM培养F3代细胞显微镜观察结果。
图2为LSM培养F3代细胞显微镜观察结果。
图3为家兔校验中家兔反应为早期(61h)出现双定性热反应。
图4为家兔校验中家兔反应为早期(60h)出现双定性热反应。
具体实施方式
下面结合具体实施方式对本发明的技术方案作进一步详细地说明。
实施例1
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：甘氨酸7.5mg/L、丙氨酸8.9mg/L、盐酸精氨酸126mg/L、一水天门冬酰胺13.2mg/L、天门冬氨酸13.3mg/L、二盐酸胱氨酸31.28mg/L、谷氨酸14.7mg/L、谷氨酰胺292mg/L、一水盐酸组氨酸42mg/L、异亮氨酸52mg/L、亮氨酸52mg/L、盐酸赖氨酸72.5mg/L、甲硫氨酸15mg/L、苯丙氨酸32mg/L、脯氨酸11.5mg/L、丝氨酸10.5mg/L、苏氨酸48mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物51.9mg/L、缬氨酸46mg/L、羟脯氨酸6.8mg/L、生物素0.15mg/L、氯化胆碱3.5mg/L、D-泛酸钙3.5mg/L、叶酸3.5mg/L、烟酰胺3.5mg/L、吡哆素盐酸盐0.2mg/L、盐酸吡哆醛0.1mg/L、核黄素0.25mg/L、盐酸硫胺素3.5mg/L、B12维生素0.2mg/L、i-肌醇6.3mg/L、维生素A醋酸酯0.05mg/L、维生素D 0.07mg/L、维生素K 0.008mg/L、无水氯化钙180mg/L、无水硫酸镁92mg/L、氯化钾380mg/L、碳酸氢钠2700mg/L、氯化钠5600mg/L、一水磷酸二氢钠130mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.3mg/L、无水氯化镁25mg/L、D-葡萄糖4000mg/L、乙醇胺2.5mg/L、次黄嘌呤钠0.4mg/L、亚油酸0.2mg/L、酚红12mg/L、腐胺二盐酸5.5mg/L、丙酮酸钠105mg/L、还原型谷胱甘肽1.7mg/L、半乳糖200mg/L、腺嘌呤12.5mg/L、血清素5.5mg/L、水溶性胆固醇2.8mg/L、盐酸鸟嘌呤14.5mg/L、胸腺嘧啶0.2mg/L、尿嘧啶0.2mg/L、胸腺嘧啶核苷0.38mg/L、人转铁蛋白6.5mg/L、重组胰岛素10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)2800mg/L。
其中，人转铁蛋白为饱和铁人转铁蛋白；重组胰岛素为全链重组胰岛素。
实施例2
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：甘氨酸5mg/L、丙氨酸5mg/L、盐酸精氨酸110mg/L、一水天门冬酰胺10mg/L、天门冬氨酸10mg/L、二盐酸胱氨酸20mg/L、谷氨酸10mg/L、谷氨酰胺200mg/L、一水盐酸组氨酸25mg/L、异亮氨酸25mg/L、亮氨酸25mg/L、盐酸赖氨酸50mg/L、甲硫氨酸10mg/L、苯丙氨酸20mg/L、脯氨酸10mg/L、丝氨酸10mg/L、苏氨酸25mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物30mg/L、缬氨酸25mg/L、羟脯氨酸5mg/L、生物素0.05mg/L、氯化胆碱2mg/L、D-泛酸钙2mg/L、叶酸2mg/L、烟酰胺2mg/L、吡哆素盐酸盐0.1mg/L、盐酸吡哆醛0.1mg/L、核黄素0.1mg/L、盐酸硫胺素1mg/L、B12维生素0.1mg/L、i-肌醇5mg/L、维生素A醋酸酯0.01mg/L、维生素D 0.01mg/L、维生素K 0.005mg/L、无水氯化钙150mg/L、无水硫酸镁50mg/L、氯化钾300mg/L、碳酸氢钠2500mg/L、氯化钠5000mg/L、一水磷酸二氢钠120mg/L、五水硫酸铜0.0005mg/L、七水合硫酸亚铁0.1mg/L、无水氯化镁10mg/L、D-葡萄糖2000mg/L、乙醇胺1mg/L、次黄嘌呤钠0.1mg/L、亚油酸0.1mg/L、酚红10mg/L、腐胺二盐酸2mg/L、丙酮酸钠90mg/L、还原型谷胱甘肽0.5mg/L、半乳糖100mg/L、腺嘌呤5mg/L、血清素5mg/L、水溶性胆固醇1mg/L、盐酸鸟嘌呤10mg/L、胸腺嘧啶0.1mg/L、尿嘧啶0.1mg/L、胸腺嘧啶核苷0.1mg/L、人转铁蛋白5mg/L、重组胰岛素5mg/L、4-羟乙基哌嗪乙磺酸(HEPES)1500mg/L。
其中，人转铁蛋白为饱和铁人转铁蛋白；重组胰岛素为全链重组胰岛素。
实施例3
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：甘氨酸10mg/L、丙氨酸10mg/L、盐酸精氨酸150mg/L、一水天门冬酰胺15mg/L、天门冬氨酸15mg/L、二盐酸胱氨酸40mg/L、谷氨酸15mg/L、谷氨酰胺350mg/L、一水盐酸组氨酸50mg/L、异亮氨酸60mg/L、亮氨酸60mg/L、盐酸赖氨酸80mg/L、甲硫氨酸20mg/L、苯丙氨酸40mg/L、脯氨酸15mg/L、丝氨酸15mg/L、苏氨酸50mg/L、色氨酸15mg/L、酪氨酸二钠盐二水合物60mg/L、缬氨酸50mg/L、羟脯氨酸10mg/L、生物素0.2mg/L、氯化胆碱5mg/L、D-泛酸钙5mg/L、叶酸5mg/L、烟酰胺5mg/L、吡哆素盐酸盐0.3mg/L、盐酸吡哆醛0.3mg/L、核黄素0.3mg/L、盐酸硫胺素5mg/L、B12维生素0.3mg/L、i-肌醇10mg/L、维生素A醋酸酯0.05mg/L、维生素D 0.1mg/L、维生素K 0.001mg/L、无水氯化钙200mg/L、无水硫酸镁120mg/L、氯化钾400mg/L、碳酸氢钠3700mg/L、氯化钠6000mg/L、一水磷酸二氢钠150mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.5mg/L、无水氯化镁30mg/L、D-葡萄糖5000mg/L、乙醇胺5mg/L、次黄嘌呤钠0.5mg/L、亚油酸0.5mg/L、酚红15mg/L、腐胺二盐酸6mg/L、丙酮酸钠120mg/L、还原型谷胱甘肽2mg/L、半乳糖300mg/L、腺嘌呤15mg/L、血清素15mg/L、水溶性胆固醇5mg/L、盐酸鸟嘌呤20mg/L、胸腺嘧啶0.5mg/L、尿嘧啶0.5mg/L、胸腺嘧啶核苷0.5mg/L、人转铁蛋白10mg/L、重组胰岛素10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)3400mg/L。
其中，人转铁蛋白为饱和铁人转铁蛋白；重组胰岛素为全链重组胰岛素。
实施例4
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：甘氨酸5mg/L、丙氨酸5mg/L、盐酸精氨酸110mg/L、一水天门冬酰胺10mg/L、天门冬氨酸10mg/L、二盐酸胱氨酸20mg/L、谷氨酸10mg/L、谷氨酰胺200mg/L、一水盐酸组氨酸25mg/L、异亮氨酸25mg/L、亮氨酸25mg/L、盐酸赖氨酸50mg/L、甲硫氨酸10mg/L、苯丙氨酸20mg/L、脯氨酸10mg/L、丝氨酸10mg/L、苏氨酸25mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物30mg/L、缬氨酸25mg/L、羟脯氨酸5mg/L、生物素0.05mg/L、氯化胆碱2mg/L、D-泛酸钙2mg/L、叶酸2mg/L、烟酰胺2mg/L、吡哆素盐酸盐0.1mg/L、盐酸吡哆醛0.1mg/L、核黄素0.1mg/L、盐酸硫胺素1mg/L、B12维生素0.1mg/L、i-肌醇5mg/L、维生素A醋酸酯0.01mg/L、维生素D 0.01mg/L、维生素K 0.005mg/L、无水氯化钙180mg/L、无水硫酸镁92mg/L、氯化钾380mg/L、碳酸氢钠2700mg/L、氯化钠5600mg/L、一水磷酸二氢钠130mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.3mg/L、无水氯化镁25mg/L、D-葡萄糖4000mg/L、乙醇胺2.5mg/L、次黄嘌呤钠0.4mg/L、亚油酸0.2mg/L、酚红12mg/L、腐胺二盐酸5.5mg/L、丙酮酸钠105mg/L、还原型谷胱甘肽1.7mg/L、半乳糖200mg/L、腺嘌呤12.5mg/L、血清素5.5mg/L、水溶性胆固醇2.8mg/L、盐酸鸟嘌呤14.5mg/L、胸腺嘧啶0.2mg/L、尿嘧啶0.2mg/L、胸腺嘧啶核苷0.38mg/L、人转铁蛋白6.5mg/L、重组胰岛素10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)2800mg/L。
其中，人转铁蛋白为饱和铁人转铁蛋白；重组胰岛素为全链重组胰岛素。
实施例5
一种用于ST细胞培养及相应病毒生产的低血清培养基，包括以下组成成分：甘氨酸7.5mg/L、丙氨酸8.9mg/L、盐酸精氨酸126mg/L、一水天门冬酰胺13.2mg/L、天门冬氨酸13.3mg/L、二盐酸胱氨酸31.28mg/L、谷氨酸14.7mg/L、谷氨酰胺292mg/L、一水盐酸组氨酸42mg/L、异亮氨酸52mg/L、亮氨酸52mg/L、盐酸赖氨酸72.5mg/L、甲硫氨酸15mg/L、苯丙氨酸32mg/L、脯氨酸11.5mg/L、丝氨酸10.5mg/L、苏氨酸48mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物51.9mg/L、缬氨酸46mg/L、羟脯氨酸6.8mg/L、生物素0.15mg/L、氯化胆碱3.5mg/L、D-泛酸钙3.5mg/L、叶酸3.5mg/L、烟酰胺3.5mg/L、吡哆素盐酸盐0.2mg/L、盐酸吡哆醛0.1mg/L、核黄素0.25mg/L、盐酸硫胺素3.5mg/L、B12维生素0.2mg/L、i-肌醇6.3mg/L、维生素A醋酸酯0.05mg/L、维生素D 0.07mg/L、维生素K 0.008mg/L、无水氯化钙180mg/L、无水硫酸镁92mg/L、氯化钾380mg/L、碳酸氢钠2700mg/L、氯化钠5600mg/L、一水磷酸二氢钠130mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.3mg/L、无水氯化镁25mg/L、D-葡萄糖5000mg/L、乙醇胺5mg/L、次黄嘌呤钠0.5mg/L、亚油酸0.5mg/L、酚红15mg/L、腐胺二盐酸6mg/L、丙酮酸钠120mg/L、还原型谷胱甘肽2mg/L、半乳糖300mg/L、腺嘌呤15mg/L、血清素15mg/L、水溶性胆固醇5mg/L、盐酸鸟嘌呤20mg/L、胸腺嘧啶0.5mg/L、尿嘧啶0.5mg/L、胸腺嘧啶核苷0.5mg/L、人转铁蛋白10mg/L、重组胰岛素10mg/L、4-羟乙基哌嗪乙磺酸(HEPES)3400mg/L。
其中，人转铁蛋白为饱和铁人转铁蛋白；重组胰岛素为全链重组胰岛素。
采用实施例1-5配方配制的培养基与对照组进行细胞培养测试：
Figure BDA0002211644710000071
实验组细胞状态良好，与阳性对照没有较大差别。而阴性对照组细胞数较少，有较多的死亡细胞。
ST细胞低血清培养体系增殖猪瘟病毒实验
Figure BDA0002211644710000081
实验步骤如下：
1)取常规条件培养良好的单层ST细胞，每克氏瓶接种约2.0×107cells，共计接种2瓶；
2)对照组添加MEM+8％BS生长液约120ml。实验组添加LSM+3％BS生长液约120ml；
3)37℃静止密闭培养72h后，以3.1方式连续传代3代；
4)分别对对照组、实验组的F3代细胞评价生长状态，并进行细胞计数。3.5将F3代生长良好的单层细胞，接入符合要求的猪瘟种毒，接毒量3％，每收次按规定时间收获病毒，共计五收次。收获后病毒将1-5收次混合，用家兔检测50万倍稀释后的RID。
实验结果
显微镜下观察可见实验组与对照组细胞生长形态均一，无聚团、单细胞巨化生长，细胞变圆界线清晰，细胞生长状态良好，二者无显著差异(图中颜色差异为显微镜光线差异)(图1、图2)
利用血球计数板将F3代实验ST细胞进行计数，结果如下(表1)：
表1.
Figure BDA0002211644710000091
计数结果显示，实验组与对照组细胞数量无显著差异。
1-5收次混合后利用家兔进行校检结果显示：利用LSM培养的ST细胞可以较好地增殖猪瘟病毒，且家兔反应为早期(60h、61h)出现双定性热反应，病毒滴度不小于50万倍RID。本次实验未按规定将每收次分别测定RID，也未进一步做100万倍RID检测，旨在验证总体产毒情况，推断部分单收次可能超过100万倍RID(图3、图4)。
本发明制备的用于ST细胞培养及相应病毒生产的低血清培养基中，使用甲骨文生物ST低血清培养基IV系列添加3％猪瘟专用新生牛血清情况下能够替代传统细胞培养基对ST细胞进行常规细胞培养，且细胞培养状态与数量均与传统培养方法无显著差异，保持细胞良好的生长状态；同时利用该培养体系可以较好地增殖猪瘟病毒，病毒滴度不小于50万倍RID。
上面对本发明的较佳实施方式作了详细说明，但是本发明并不限于上述实施方式，在本领域的普通技术人员所具备的知识范围内，还可以在不脱离本发明宗旨的前提下作出各种变化。

Claims (7)

1.一种用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，包括以下组成成分：氨基酸、维生素、无机盐类、辅助成分和蛋白质；所述氨基酸包括甘氨酸5-10mg/L、丙氨酸5-10mg/L、盐酸精氨110-150mg/L、一水天门冬酰10-15mg/L、天门冬氨10-15mg/L、二盐酸胱氨酸20-40mg/L、谷氨酸10-15mg/L、谷氨酰胺200-350mg/L、一水盐酸组氨酸25-50mg/L、异亮氨酸25-60mg/L、亮氨酸25-60mg/L、盐酸赖氨酸50-80mg/L、甲硫氨酸10-20mg/L、苯丙氨酸20-40mg/L、脯氨酸10-15mg/L、丝氨酸10-15mg/L、苏氨酸25-50mg/L、色氨酸 5-15mg/L、酪氨酸二钠盐二水合物30-60mg/L、缬氨酸25-50mg/L、羟脯氨酸5-10mg/L；所述蛋白质包括人转铁蛋白5-10mg/L、重组胰岛素5-10mg/L、4-羟乙基哌嗪乙磺酸 1500-3400mg/L。
2.根据权利要求1所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，所述维生素包括生物素0.05-0.2mg/L、氯化胆碱2-5mg/L、D-泛酸钙 2-5mg/L、叶酸2-5mg/L、烟酰胺2-5mg/L、吡哆素盐酸盐0.1-0.3mg/L、盐酸吡哆醛0.1-0.3mg/L、核黄素0.1-0.3mg/L、盐酸硫胺素1-5mg/L、B12维生素 0.1-0.3mg/L、i-肌醇5-10mg/L、维生素A醋酸酯0.01-0.05mg/L、维生素D 0.01-0.1mg/L、维生素K 0.005-0.01mg/L。
3.根据权利要求2所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，所述无机盐类包括无水氯化钙 150-200mg/L、无水硫酸镁 50-120mg/L、氯化钾300-400mg/L、碳酸氢钠2500-3700mg/L、氯化钠5000-6000mg/L、一水磷酸二氢钠120-150mg/L、五水硫酸铜0.0005-0.001mg/L、七水合硫酸亚铁0.1-0.5mg/L、无水氯化镁10-30mg/L。
4.根据权利要求3所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，所述辅助成分包括D-葡萄糖2000-5000mg/L、乙醇胺1-5mg/L、次黄嘌呤钠0.1-0.5mg/L、亚油酸0.1-0.5mg/L、酚红10-15mg/L、腐胺二盐酸 2-6mg/L、丙酮酸钠90-120mg/L、还原型谷胱甘肽0.5-2.0mg/L、半乳糖 100-300mg/L、腺嘌呤5-15mg/L、血清素5-15mg/L、水溶性胆固醇1-5mg/L、盐酸鸟嘌呤10-20mg/L、胸腺嘧啶0.1-0.5mg/L、尿嘧啶0.1-0.5mg/L、胸腺嘧啶核苷0.1-0.5mg/L。
5.根据权利要求4所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，包括以下组成成分：甘氨酸7.5mg/L、丙氨酸8.9mg/L、盐酸精氨酸126mg/L 、一水天门冬酰胺13.2mg/L 、天门冬氨酸13.3mg/L 、二盐酸胱氨酸31.28mg/L、谷氨酸14.7mg/L、谷氨酰胺292mg/L、一水盐酸组氨酸 42mg/L、异亮氨酸52mg/L、亮氨酸52mg/L、盐酸赖氨酸72.5mg/L、甲硫氨酸15mg/L、苯丙氨酸 32mg/L 、脯氨酸11.5mg/L、丝氨酸10.5mg/L、苏氨酸48mg/L、色氨酸10mg/L、酪氨酸二钠盐二水合物 51.9mg/L、缬氨酸46mg/L 、羟脯氨酸 6.8mg/L、生物素0.15mg/L、氯化胆碱3.5mg/L、D-泛酸钙3.5mg/L、叶酸3.5mg/L、烟酰胺3.5mg/L、吡哆素盐酸盐0.2mg/L 、盐酸吡哆醛0.1mg/L、核黄素0.25mg/L 、盐酸硫胺素3.5mg/L、B12维生素0.2mg/L、i-肌醇 6.3mg/L、维生素A醋酸酯0.05mg/L 、维生素D 0.07mg/L 、维生素K0.008mg/L、无水氯化钙180mg/L、无水硫酸镁92mg/L、氯化钾380mg/L、碳酸氢钠2700mg/L、氯化钠5600mg/L、一水磷酸二氢钠130mg/L、五水硫酸铜0.001mg/L、七水合硫酸亚铁0.3mg/L、无水氯化镁25mg/L、D-葡萄糖4000mg/L、乙醇胺2.5mg/L、次黄嘌呤钠 0.4mg/L、亚油酸0.2mg/L、酚红12mg/L、腐胺二盐酸5.5mg/L 、丙酮酸钠105mg/L 、还原型谷胱甘肽1.7mg/L、半乳糖200mg/L、腺嘌呤12.5mg/L、血清素 5.5mg/L、水溶性胆固醇2.8mg/L、盐酸鸟嘌呤14.5mg/L、胸腺嘧啶0.2mg/L、尿嘧啶0.2mg/L、胸腺嘧啶核苷0.38mg/L、人转铁蛋白6.5mg/L、重组胰岛素 10mg/L、4-羟乙基哌嗪乙磺酸 2800mg/L。
6.根据权利要求5所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，所述人转铁蛋白为饱和铁人转铁蛋白。
7.根据权利要求5所述的用于ST细胞培养及相应病毒生产的低血清培养基，其特征在于，所述重组胰岛素为全链重组胰岛素。
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