CN112458052A - 一种免疫细胞培养基 - Google Patents

一种免疫细胞培养基 Download PDF

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CN112458052A
CN112458052A CN202011437654.2A CN202011437654A CN112458052A CN 112458052 A CN112458052 A CN 112458052A CN 202011437654 A CN202011437654 A CN 202011437654A CN 112458052 A CN112458052 A CN 112458052A
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高旭
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Tianjin Haoyang Biological Products Technology Co ltd
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Abstract

本发明提出一种免疫细胞培养基，包括如下组份：氨基酸混合物、氨基酸盐混合物、酰胺混合物、无机盐混合物、D‑泛酸钙、氯化胆碱、盐酸吡哆醛、盐酸硫胺素、肌醇、生物素、维生素、叶酸、核黄素、丙酮酸钠、葡萄糖、酚红、4‑羟乙基哌嗪乙磺酸、人血白蛋白、转铁蛋白、维生素C磷酸镁、肝素钠、盐酸腐胺、胰岛素、超氧化物歧化酶、胆固醇、油酸、生育酚、棕榈酸、维生素A醋酸酯、碳酸氢钠和甲萘醌。本发明不用补充自体血浆即可适合免疫细胞快速增殖，显著缩短了免疫细胞的培养周期，二维、三维的细胞培养体系均适用，应用范围更广。

Description

一种免疫细胞培养基
技术领域
本发明属于细胞生物学技术领域，特别是指一种免疫细胞培养基。
背景技术
无血清培养基是指不需要添加血清就可以维持细胞在体外较长时间生长繁殖的合成培养基，可以应用于培养免疫细胞，例如淋巴细胞、树突状细胞、单核/巨噬细胞等。与有血清培养基相比，无血清培养基不含对细胞有害的动物成分，而且价格相对低。然而，现有的无血清培养基培养免疫细胞时，为了加速细胞繁殖，缩短培养时间，在细胞培养前期需要补充自体血浆，否则很难达到临床的回输标准，而且所耗费的培养时间也更长，在三维培养体系中，该缺陷尤为显著，即使补充自体血浆，免疫细胞的增殖速度也比较慢，所需的培养时间远大于有血清培养基。因此，研发一种不用补充自体血浆即可适合免疫细胞快速增殖、有效缩短培养周期的免疫细胞培养基，是目前急需解决的技术问题。
发明内容
为解决以上现有技术的不足，本发明提出了一种适合免疫细胞快速增殖、有效缩短培养周期的免疫细胞培养基。
本发明的技术方案是这样实现的：
一种免疫细胞培养基，包括如下组份：氨基酸混合物、氨基酸盐混合物、酰胺混合物、无机盐混合物、D-泛酸钙、氯化胆碱、盐酸吡哆醛、盐酸硫胺素、肌醇、生物素、维生素、叶酸、核黄素、丙酮酸钠、葡萄糖、酚红、4-羟乙基哌嗪乙磺酸、人血白蛋白、转铁蛋白、维生素C磷酸镁、肝素钠、盐酸腐胺、胰岛素、超氧化物歧化酶、胆固醇、油酸、生育酚、棕榈酸、维生素A醋酸酯、碳酸氢钠和甲萘醌。
经过大量实验研究发现：上述组份以特定的浓度进行配比，各个组份互相协调、共同作用，提供免疫细胞增殖所需要的充足的营养和良好的环境，可以显著提高免疫细胞的增殖速度，极大缩短免疫细胞的培养时间；而且，令人意外的是，本发明在促进免疫细胞高速繁殖的同时，还显著提高了免疫细胞的存活率，而且临床安全性好。
优选的，氨基酸混合物由甘氨酸、L-丙氨酸、L-天冬氨酸、L-谷氨酸、盐酸组氨酸、异亮氨酸、L-亮氨酸、L-蛋氨酸、L-苯丙氨酸、L-脯氨酸、L-丝氨酸、苏氨酸、L-色氨酸和L-缬氨酸组成；氨基酸盐混合物由L-精氨酸盐酸盐、L-胱氨酸二盐酸盐、赖氨酸盐酸盐和L-酪氨酸二钠组成；酰胺混合物由L-天冬酰胺、L-谷氨酰胺和烟酰胺组成；无机盐混合物由磷酸二氢钠、氯化钙、氯化钠、硫酸镁、氯化钾、亚硒酸钠、五水硫酸铜和九水合硝酸铁组成。
更为优选的，各个组份的含量分别为：甘氨酸25-35mg/L、L-丙氨酸20-30mg/L、L-精氨酸盐酸盐65-100mg/L、L-天冬酰胺20-30mg/L、L-天冬氨酸25-35mg/L、L-胱氨酸二盐酸盐80-100mg/L、L-谷氨酸60-85mg/L、L-谷氨酰胺500-650mg/L、盐酸组氨酸30-50mg/L、异亮氨酸80-120mg/L、L-亮氨酸80-120mg/L、赖氨酸盐酸盐130-160mg/L、L-蛋氨酸25-35mg/L、L-苯丙氨酸50-80mg/L、L-脯氨酸30-50mg/L、L-丝氨酸35-50mg/L、苏氨酸80-110mg/L、L-色氨酸10-20mg/L、L-酪氨酸二钠80-120mg/L、L-缬氨酸80-110mg/L、磷酸二氢钠100-150mg/L、D-泛酸钙8-12mg/L、氯化钙100-130mg/L、氯化钠3500-6000mg/L、硫酸镁60-130mg/L、氯化胆碱10-14mg/L、烟酰胺10-14mg/L、盐酸吡哆醛10-14mg/L、盐酸硫胺素10-14mg/L、肌醇1-3mg/L、生物素1-3mg/L、维生素B12 1-3mg/L、叶酸1-3mg/L、核黄素1-3mg/L、丙酮酸钠90-130mg/L、氯化钾25-35mg/L、葡萄糖4000-5000mg/L、酚红15-22mg/L、4-羟乙基哌嗪乙磺酸1800-2200mg/L、亚硒酸钠0.01-0.02mg/L、五水硫酸铜0.0005-0.0005mg/L、九水合硝酸铁0.0005-0.005mg/L、人血白蛋白4000-6000mg/L、转铁蛋白15-40mg/L、维生素C磷酸镁400-600mg/L、肝素钠5-8mg/L、盐酸腐胺5-8mg/L、胰岛素20-30mg/L、超氧化物歧化酶30-50mg/L、胆固醇10-14mg/L、油酸2-6mg/L、生育酚1-5mg/L、棕榈酸1-5mg/L、维生素A醋酸酯1-5mg/L、碳酸氢钠2500-3500mg/L、甲萘醌0.005-0.015mg/L。
最为优选的，各个组份的含量分别为：甘氨酸30mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐84mg/L、L-天冬酰胺25mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸75mg/L、L-谷氨酰胺584mg/L、盐酸组氨酸42mg/L、异亮氨酸105mg/L、L-亮氨酸105mg/L、赖氨酸盐酸盐146mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸66mg/L、L-脯氨酸40mg/L、L-丝氨酸42mg/L、苏氨酸95mg/L、L-色氨酸16mg/L、L-酪氨酸二钠104mg/L、L-缬氨酸94mg/L、磷酸二氢钠125mg/L、D-泛酸钙8-12mg/L、氯化钙117mg/L、氯化钠4500mg/L、硫酸镁98mg/L、氯化胆碱10-14mg/L、烟酰胺10-14mg/L、盐酸吡哆醛10-14mg/L、盐酸硫胺素10-14mg/L、肌醇1-3mg/L、生物素1-3mg/L、维生素B12 1-3mg/L、叶酸1-3mg/L、核黄素1-3mg/L、丙酮酸钠110mg/L、氯化钾30mg/L、葡萄糖4500mg/L、酚红20mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.017mg/L、五水硫酸铜0.001mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白15-40mg/L、维生素C磷酸镁500mg/L、肝素钠5-8mg/L、盐酸腐胺5-8mg/L、胰岛素25mg/L、超氧化物歧化酶40mg/L、胆固醇10-14mg/L、油酸2-6mg/L、生育酚3mg/L、棕榈酸3mg/L、维生素A醋酸酯3mg/L、碳酸氢钠3025mg/L、甲萘醌0.01mg/L。
与现有的无血清免疫细胞培养基相比，本发明不用补充自体血浆即可适合免疫细胞快速增殖，显著缩短了免疫细胞的培养周期，二维、三维的细胞培养体系均适用，应用范围更广。
具体实施方式
下面将结合本发明实施例，对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
以下实施例所用的原料均通过商购获得。配制时，先在容器内加入略少于培养基总体积的水，室温下向水中加入除去碳酸氢钠之外的其他各组份并轻轻搅拌至均匀，之后加入碳酸氢钠，搅拌溶解后定容并过滤无菌、分装、封口，于4℃下冷藏备用。此处所用的水至少为双蒸水，优选三蒸水。
实施例1
一种免疫细胞培养基，包括如下浓度的组份：甘氨酸35mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐65mg/L、L-天冬酰胺30mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐80mg/L、L-谷氨酸85mg/L、L-谷氨酰胺580mg/L、盐酸组氨酸30mg/L、异亮氨酸120mg/L、L-亮氨酸100mg/L、赖氨酸盐酸盐130mg/L、L-蛋氨酸35mg/L、L-苯丙氨酸60mg/L、L-脯氨酸30mg/L、L-丝氨酸50mg/L、苏氨酸90mg/L、L-色氨酸10mg/L、L-酪氨酸二钠120mg/L、L-缬氨酸90mg/L、磷酸二氢钠100mg/L、D-泛酸钙12mg/L、氯化钙120mg/L、氯化钠3500mg/L、硫酸镁130mg/L、氯化胆碱12mg/L、烟酰胺10mg/L、盐酸吡哆醛14mg/L、盐酸硫胺素12mg/L、肌醇1mg/L、生物素3mg/L、维生素B12 2mg/L、叶酸1mg/L、核黄素3mg/L、丙酮酸钠110mg/L、氯化钾25mg/L、葡萄糖5000mg/L、酚红18mg/L、4-羟乙基哌嗪乙磺酸1800mg/L、亚硒酸钠0.02mg/L、五水硫酸铜0.0001mg/L、九水合硝酸铁0.0005mg/L、人血白蛋白6000mg/L、转铁蛋白30mg/L、维生素C磷酸镁400mg/L、肝素钠8mg/L、盐酸腐胺6.5mg/L、胰岛素20mg/L、超氧化物歧化酶50mg/L、胆固醇12mg/L、油酸2mg/L、生育酚5mg/L、棕榈酸3mg/L、维生素A醋酸酯1mg/L、碳酸氢钠3500mg/L、甲萘醌0.01mg/L，其余为水。
实施例2
一种免疫细胞培养基，包括如下组份：甘氨酸25mg/L、L-丙氨酸30mg/L、L-精氨酸盐酸盐80mg/L、L-天冬酰胺20mg/L、L-天冬氨酸35mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸60mg/L、L-谷氨酰胺650mg/L、盐酸组氨酸40mg/L、异亮氨酸80mg/L、L-亮氨酸120mg/L、赖氨酸盐酸盐150mg/L、L-蛋氨酸25mg/L、L-苯丙氨酸80mg/L、L-脯氨酸40mg/L、L-丝氨酸35mg/L、苏氨酸110mg/L、L-色氨酸15mg/L、L-酪氨酸二钠80mg/L、L-缬氨酸110mg/L、磷酸二氢钠130mg/L、D-泛酸钙8mg/L、氯化钙130mg/L、氯化钠4500mg/L、硫酸镁60mg/L、氯化胆碱14mg/L、烟酰胺12mg/L、盐酸吡哆醛10mg/L、盐酸硫胺素14mg/L、肌醇2mg/L、生物素1mg/L、维生素B12 3mg/L、叶酸2mg/L、核黄素1mg/L、丙酮酸钠130mg/L、氯化钾30mg/L、葡萄糖4000mg/L、酚红22mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.01mg/L、五水硫酸铜0.0005mg/L、九水合硝酸铁0.003mg/L、人血白蛋白4000mg/L、转铁蛋白40mg/L、维生素C磷酸镁500mg/L、肝素钠5mg/L、盐酸腐胺8mg/L、胰岛素25mg/L、超氧化物歧化酶30mg/L、胆固醇14mg/L、油酸4mg/L、生育酚1mg/L、棕榈酸5mg/L、维生素A醋酸酯3mg/L、碳酸氢钠2500mg/L、甲萘醌0.015mg/L，其余为水。
实施例3
一种免疫细胞培养基，包括如下组份：甘氨酸30mg/L、L-丙氨酸20mg/L、L-精氨酸盐酸盐100mg/L、L-天冬酰胺25mg/L、L-天冬氨酸25mg/L、L-胱氨酸二盐酸盐100mg/L、L-谷氨酸75mg/L、L-谷氨酰胺500mg/L、盐酸组氨酸50mg/L、异亮氨酸100mg/L、L-亮氨酸80mg/L、赖氨酸盐酸盐160mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸50mg/L、L-脯氨酸50mg/L、L-丝氨酸45mg/L、苏氨酸80mg/L、L-色氨酸20mg/L、L-酪氨酸二钠100mg/L、L-缬氨酸80mg/L、磷酸二氢钠150mg/L、D-泛酸钙10mg/L、氯化钙100mg/L、氯化钠6000mg/L、硫酸镁100mg/L、氯化胆碱10mg/L、烟酰胺14mg/L、盐酸吡哆醛12mg/L、盐酸硫胺素10mg/L、肌醇3mg/L、生物素2mg/L、维生素B12 1mg/L、叶酸3mg/L、核黄素2mg/L、丙酮酸钠90mg/L、氯化钾35mg/L、葡萄糖4500mg/L、酚红15mg/L、4-羟乙基哌嗪乙磺酸2200mg/L、亚硒酸钠0.015mg/L、五水硫酸铜0.0005mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白15mg/L、维生素C磷酸镁600mg/L、肝素钠7mg/L、盐酸腐胺5mg/L、胰岛素30mg/L、超氧化物歧化酶40mg/L、胆固醇10mg/L、油酸6mg/L、生育酚4mg/L、棕榈酸1mg/L、维生素A醋酸酯5mg/L、碳酸氢钠2800mg/L、甲萘醌0.005mg/L，其余为水。
实施例4
一种免疫细胞培养基，包括如下组份：甘氨酸30mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐84mg/L、L-天冬酰胺25mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸75mg/L、L-谷氨酰胺584mg/L、盐酸组氨酸42mg/L、异亮氨酸105mg/L、L-亮氨酸105mg/L、赖氨酸盐酸盐146mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸66mg/L、L-脯氨酸40mg/L、L-丝氨酸42mg/L、苏氨酸95mg/L、L-色氨酸16mg/L、L-酪氨酸二钠104mg/L、L-缬氨酸94mg/L、磷酸二氢钠125mg/L、D-泛酸钙12mg/L、氯化钙117mg/L、氯化钠4500mg/L、硫酸镁98mg/L、氯化胆碱12mg/L、烟酰胺10mg/L、盐酸吡哆醛14mg/L、盐酸硫胺素12mg/L、肌醇1mg/L、生物素3mg/L、维生素B12 2mg/L、叶酸1mg/L、核黄素3mg/L、丙酮酸钠110mg/L、氯化钾30mg/L、葡萄糖4500mg/L、酚红20mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.017mg/L、五水硫酸铜0.001mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白30mg/L、维生素C磷酸镁500mg/L、肝素钠5mg/L、盐酸腐胺5-8mg/L、胰岛素25mg/L、超氧化物歧化酶40mg/L、胆固醇14mg/L、油酸4mg/L、生育酚3mg/L、棕榈酸3mg/L、维生素A醋酸酯3mg/L、碳酸氢钠3025mg/L、甲萘醌0.01mg/L，其余为水。
实施例5
一种免疫细胞培养基，包括如下组份：甘氨酸30mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐84mg/L、L-天冬酰胺25mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸75mg/L、L-谷氨酰胺584mg/L、盐酸组氨酸42mg/L、异亮氨酸105mg/L、L-亮氨酸105mg/L、赖氨酸盐酸盐146mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸66mg/L、L-脯氨酸40mg/L、L-丝氨酸42mg/L、苏氨酸95mg/L、L-色氨酸16mg/L、L-酪氨酸二钠104mg/L、L-缬氨酸94mg/L、磷酸二氢钠125mg/L、D-泛酸钙8mg/L、氯化钙117mg/L、氯化钠4500mg/L、硫酸镁98mg/L、氯化胆碱14mg/L、烟酰胺13mg/L、盐酸吡哆醛10mg/L、盐酸硫胺素14mg/L、肌醇2mg/L、生物素1mg/L、维生素B12 3mg/L、叶酸2mg/L、核黄素1mg/L、丙酮酸钠110mg/L、氯化钾30mg/L、葡萄糖4500mg/L、酚红20mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.017mg/L、五水硫酸铜0.001mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白40mg/L、维生素C磷酸镁500mg/L、肝素钠7mg/L、盐酸腐胺5mg/L、胰岛素25mg/L、超氧化物歧化酶40mg/L、胆固醇14mg/L、油酸3mg/L、生育酚3mg/L、棕榈酸3mg/L、维生素A醋酸酯3mg/L、碳酸氢钠3025mg/L、甲萘醌0.01mg/L，其余为水。
实施例6
一种免疫细胞培养基，包括如下组份：甘氨酸30mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐84mg/L、L-天冬酰胺25mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸75mg/L、L-谷氨酰胺584mg/L、盐酸组氨酸42mg/L、异亮氨酸105mg/L、L-亮氨酸105mg/L、赖氨酸盐酸盐146mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸66mg/L、L-脯氨酸40mg/L、L-丝氨酸42mg/L、苏氨酸95mg/L、L-色氨酸16mg/L、L-酪氨酸二钠104mg/L、L-缬氨酸94mg/L、磷酸二氢钠125mg/L、D-泛酸钙10mg/L、氯化钙117mg/L、氯化钠4500mg/L、硫酸镁98mg/L、氯化胆碱10mg/L、烟酰胺14mg/L、盐酸吡哆醛13mg/L、盐酸硫胺素10mg/L、肌醇3mg/L、生物素2mg/L、维生素B12 1mg/L、叶酸3mg/L、核黄素2mg/L、丙酮酸钠110mg/L、氯化钾30mg/L、葡萄糖4500mg/L、酚红20mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.017mg/L、五水硫酸铜0.001mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白15mg/L、维生素C磷酸镁500mg/L、肝素钠8mg/L、盐酸腐胺7mg/L、胰岛素25mg/L、超氧化物歧化酶40mg/L、胆固醇10mg/L、油酸6mg/L、生育酚3mg/L、棕榈酸3mg/L、维生素A醋酸酯3mg/L、碳酸氢钠3025mg/L、甲萘醌0.01mg/L，其余为水。
测试表征：
一、T细胞扩增试验
以DEME(对比例1)及在售的友康(NC0101)常规RPMI1640免疫细胞无血清培养基(对比例2)作为对比，测试本发明实施例1-6所得的免疫细胞培养基的培养效果。试验时，以培养皿为容器，选用来源于人的T细胞作为培养细胞，按照104个/ml接种后分别加入到DEME、RPMI1640及本发明实施例1-6所得的产品中于37℃、5％CO2下进行培养，在培养1、7和14天分别取样进行细胞浓度的检测，并在第14天检测细胞存活率。检测结果如下表1所示：
对比例1 对比例2 实施例1 实施例2 实施例3 实施例4 实施例5 实施例6
第1天 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup>
第7天 1×10<sup>5</sup> 1.9×10<sup>5</sup> 2.5×10<sup>5</sup> 3.4×10<sup>5</sup> 2.8×10<sup>5</sup> 6.8×10<sup>5</sup> 7.8×10<sup>5</sup> 8.4×10<sup>5</sup>
第14天 2.5×10<sup>6</sup> 3.2×10<sup>6</sup> 1.8×10<sup>7</sup> 2.3×10<sup>7</sup> 2.2×10<sup>7</sup> 5.9×10<sup>7</sup> 8.5×10<sup>7</sup> 9.8×10<sup>7</sup>
存活率 85％ 89％ 96.8％ 97.3％ 96.4％ 99.1％ 98.5％ 98.2％
由表1可知：与现有技术相比，本发明所制得的免疫细胞培养基无论是细胞增殖速率，还是细胞存活率，都显著优于DEME(对比例1)及友康(NC0101)常规RPMI1640免疫细胞无血清培养基(对比例2)。
以本发明实施例6所得的产品为代表，测试其与在售的友康(NC0101)常规RPMI1640免疫细胞无血清培养基在二维培养体系与三维培养体系中的适用性。以实施例6的产品为培养基、以培养板为培养容器标记为实施例7，以实施例6的产品为培养基、以培养瓶为培养容器标记为实施例8，以RPMI1640为培养基、以培养板为培养容器标记为对比例3，以RPMI1640为培养基、以培养瓶为培养容器标记为对比例4。分别选用来源于人的T细胞作为培养细胞，按照104接种后分别加入到DEME、RPMI1640及本发明实施例1-6所得的产品中于37℃、5％CO2下进行培养，在培养1、7和14天分别取样进行细胞浓度的检测，并在第14天检测细胞存活率。检测结果如下表2所示：
对比例3 对比例4 实施例7 实施例8
第1天 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup> 10<sup>4</sup>
第7天 1.1×10<sup>5</sup> 5×10<sup>4</sup> 8.3×10<sup>5</sup> 3.1×10<sup>5</sup>
第14天 2.6×10<sup>6</sup> 1.2×10<sup>6</sup> 9.6×10<sup>7</sup> 6.9×10<sup>7</sup>
存活率 83％ 71％ 98.8％ 96.3％
由表2可知：RPMI1640用于三维培养体系时，无论是细胞增殖速率还是细胞存活率，均大幅下降；而本发明实施例6的产品用于二维培养体系(培养板等)与三维培养体系(高密度培养桶)时，所能达到的细胞增殖速率及细胞存活率基本没有很大的差别，均可以保持很高的增殖率和存活率，可见其在二维培养体系与三维培养体系中皆有特别良好的适应性。
二、NK细胞扩增试验
以美国Corning NK细胞活化/扩增培养基(产品货号：88-571-kit)作为对比培养基，测试本发明实施例1-6所得的免疫细胞培养基的培养效果。试验时，以培养皿为容器，选用NK细胞作为培养细胞，按照3×106个/ml接种后分别加入到对比培养基及本发明实施例1-6所得的产品中于饱和湿度、37℃、5％CO2培养箱中培养，在培养的第9、10、12、13、14天分别取样检测NK细胞的个数。检测结果如下表2所示：
对比培养基 实施例1 实施例2 实施例3 实施例4 实施例5 实施例6
第9天 9亿个 15.3亿个 16亿个 14.8亿个 15亿个 15亿个 14.5亿个
第10天 15亿个 28.3亿个 29亿个 27.5亿个 28亿个 27.6亿个 27亿个
第11天 28亿个 50亿个 51亿个 49亿个 49亿个 48亿个 49亿个
第12天 36亿个 61亿个 64亿个 62亿个 62亿个 63亿个 60亿个
第13天 45亿个 78亿个 79亿个 78亿个 77亿个 73亿个 75亿个
第14天 53亿个 82亿个 84亿个 80亿个 81亿个 78亿个 79亿个
经过14天的培养，采用本发明的培养基所得的NK细胞在80亿个左右，而采用对比培养基仅获得53亿个。本发明提出的免疫细胞培养基在应用于NK细胞扩增试验时，增殖率较美国Corning NK细胞活化/扩增培养基提高了至少50.9％。
以上所述仅为本发明的较佳实施例而已，其中所用的“左”、“右”等方位词皆为便于阐述本发明的结构，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (4)

1.一种免疫细胞培养基，其特征在于：包括如下组份：氨基酸混合物、氨基酸盐混合物、酰胺混合物、无机盐混合物、D-泛酸钙、氯化胆碱、盐酸吡哆醛、盐酸硫胺素、肌醇、生物素、维生素、叶酸、核黄素、丙酮酸钠、葡萄糖、酚红、4-羟乙基哌嗪乙磺酸、人血白蛋白、转铁蛋白、维生素C磷酸镁、肝素钠、盐酸腐胺、胰岛素、超氧化物歧化酶、胆固醇、油酸、生育酚、棕榈酸、维生素A醋酸酯、碳酸氢钠和甲萘醌。
2.根据权利要求1所述的免疫细胞培养基，其特征在于：所述氨基酸混合物由甘氨酸、L-丙氨酸、L-天冬氨酸、L-谷氨酸、盐酸组氨酸、异亮氨酸、L-亮氨酸、L-蛋氨酸、L-苯丙氨酸、L-脯氨酸、L-丝氨酸、苏氨酸、L-色氨酸和L-缬氨酸组成；所述氨基酸盐混合物由L-精氨酸盐酸盐、L-胱氨酸二盐酸盐、赖氨酸盐酸盐和L-酪氨酸二钠组成；所述酰胺混合物由L-天冬酰胺、L-谷氨酰胺和烟酰胺组成；所述无机盐混合物由磷酸二氢钠、氯化钙、氯化钠、硫酸镁、氯化钾、亚硒酸钠、五水硫酸铜和九水合硝酸铁组成。
3.根据权利要求2所述的免疫细胞培养基，其特征在于：各个组份的含量分别为：甘氨酸25-35mg/L、L-丙氨酸20-30mg/L、L-精氨酸盐酸盐65-100mg/L、L-天冬酰胺20-30mg/L、L-天冬氨酸25-35mg/L、L-胱氨酸二盐酸盐80-100mg/L、L-谷氨酸60-85mg/L、L-谷氨酰胺500-650mg/L、盐酸组氨酸30-50mg/L、异亮氨酸80-120mg/L、L-亮氨酸80-120mg/L、赖氨酸盐酸盐130-160mg/L、L-蛋氨酸25-35mg/L、L-苯丙氨酸50-80mg/L、L-脯氨酸30-50mg/L、L-丝氨酸35-50mg/L、苏氨酸80-110mg/L、L-色氨酸10-20mg/L、L-酪氨酸二钠80-120mg/L、L-缬氨酸80-110mg/L、磷酸二氢钠100-150mg/L、D-泛酸钙8-12mg/L、氯化钙100-130mg/L、氯化钠3500-6000mg/L、硫酸镁60-130mg/L、氯化胆碱10-14mg/L、烟酰胺10-14mg/L、盐酸吡哆醛10-14mg/L、盐酸硫胺素10-14mg/L、肌醇1-3mg/L、生物素1-3mg/L、维生素B12 1-3mg/L、叶酸1-3mg/L、核黄素1-3mg/L、丙酮酸钠90-130mg/L、氯化钾25-35mg/L、葡萄糖4000-5000mg/L、酚红15-22mg/L、4-羟乙基哌嗪乙磺酸1800-2200mg/L、亚硒酸钠0.01-0.02mg/L、五水硫酸铜0.0005-0.0005mg/L、九水合硝酸铁0.0005-0.005mg/L、人血白蛋白4000-6000mg/L、转铁蛋白15-40mg/L、维生素C磷酸镁400-600mg/L、肝素钠5-8mg/L、盐酸腐胺5-8mg/L、胰岛素20-30mg/L、超氧化物歧化酶30-50mg/L、胆固醇10-14mg/L、油酸2-6mg/L、生育酚1-5mg/L、棕榈酸1-5mg/L、维生素A醋酸酯1-5mg/L、碳酸氢钠2500-3500mg/L、甲萘醌0.005-0.015mg/L。
4.根据权利要求1所述的免疫细胞培养基，其特征在于：各个组份的含量分别为：甘氨酸30mg/L、L-丙氨酸25mg/L、L-精氨酸盐酸盐84mg/L、L-天冬酰胺25mg/L、L-天冬氨酸30mg/L、L-胱氨酸二盐酸盐90mg/L、L-谷氨酸75mg/L、L-谷氨酰胺584mg/L、盐酸组氨酸42mg/L、异亮氨酸105mg/L、L-亮氨酸105mg/L、赖氨酸盐酸盐146mg/L、L-蛋氨酸30mg/L、L-苯丙氨酸66mg/L、L-脯氨酸40mg/L、L-丝氨酸42mg/L、苏氨酸95mg/L、L-色氨酸16mg/L、L-酪氨酸二钠104mg/L、L-缬氨酸94mg/L、磷酸二氢钠125mg/L、D-泛酸钙8-12mg/L、氯化钙117mg/L、氯化钠4500mg/L、硫酸镁98mg/L、氯化胆碱10-14mg/L、烟酰胺10-14mg/L、盐酸吡哆醛10-14mg/L、盐酸硫胺素10-14mg/L、肌醇1-3mg/L、生物素1-3mg/L、维生素B12 1-3mg/L、叶酸1-3mg/L、核黄素1-3mg/L、丙酮酸钠110mg/L、氯化钾30mg/L、葡萄糖4500mg/L、酚红20mg/L、4-羟乙基哌嗪乙磺酸2000mg/L、亚硒酸钠0.017mg/L、五水硫酸铜0.001mg/L、九水合硝酸铁0.005mg/L、人血白蛋白5000mg/L、转铁蛋白15-40mg/L、维生素C磷酸镁500mg/L、肝素钠5-8mg/L、盐酸腐胺5-8mg/L、胰岛素25mg/L、超氧化物歧化酶40mg/L、胆固醇10-14mg/L、油酸2-6mg/L、生育酚3mg/L、棕榈酸3mg/L、维生素A醋酸酯3mg/L、碳酸氢钠3025mg/L、甲萘醌0.01mg/L。
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114134111A (zh) * 2021-12-07 2022-03-04 珠海贝索细胞科学技术有限公司 一种免疫细胞活化剂及其制备方法和应用
CN117546837A (zh) * 2023-11-22 2024-02-13 深圳泽医细胞治疗集团有限公司 适用于nk细胞培养的血液保存液及其制备方法和应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105039253A (zh) * 2015-08-05 2015-11-11 广州赛莱拉干细胞科技股份有限公司 适用于免疫细胞大规模培养的无血清培养基
CN108060128A (zh) * 2017-12-12 2018-05-22 成都源泉生物科技有限公司 免疫细胞用基础培养基的生产方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105039253A (zh) * 2015-08-05 2015-11-11 广州赛莱拉干细胞科技股份有限公司 适用于免疫细胞大规模培养的无血清培养基
CN108060128A (zh) * 2017-12-12 2018-05-22 成都源泉生物科技有限公司 免疫细胞用基础培养基的生产方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
朱诗国 等: "《医学免疫学》", 30 April 2020, 上海科学技术出版社 *
申春一 等: "TWS119 联合细胞因子促进CD8+记忆性T细胞分化及功能", 《中国免疫学杂志》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114134111A (zh) * 2021-12-07 2022-03-04 珠海贝索细胞科学技术有限公司 一种免疫细胞活化剂及其制备方法和应用
CN114134111B (zh) * 2021-12-07 2023-08-18 珠海贝索细胞科学技术有限公司 一种免疫细胞活化剂及其制备方法和应用
CN117546837A (zh) * 2023-11-22 2024-02-13 深圳泽医细胞治疗集团有限公司 适用于nk细胞培养的血液保存液及其制备方法和应用
CN117546837B (zh) * 2023-11-22 2024-09-03 深圳泽医细胞治疗集团有限公司 适用于nk细胞培养的血液保存液及其制备方法和应用

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