AUTOMATED LAK CELL GENERATION USING CELL SEPARATOR TECHNIQUES

使用细胞分离器技术自动生成 LAK 细胞

• 批准号: 3874309
• 负责人: S F LEITMAN
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874309
• 关键词: biomedical automation; biotechnology; blood banks; cell sorting; clinical biomedical equipment; density gradient ultracentrifugation; human subject; interleukin 2; killer cells; leukapheresis; lymphocyte; neoplasm /cancer immunotherapy; neoplasm /cancer therapy; passive immunization; tissue /cell culture

In order to simplify the production of lymphokine-activated killer (LAK) cells for therapeutic use, an automated cell separator (CS-3000, Fenwal) was adapted to: (1) process large volumes of whole blood for collection of leukocyte concentrates from cancer patients, (2) purify the lymphocyte fraction by automated Ficoll-Hypaque density gradient sedimentation, (3) culture the lymphocytes in platelet storage bags, and (4) harvest and concentrate the LAK cells out of culture. Nearly 100 LAK cell preparations have been processed in this manner. Automated density gradient sedimentation resulted in high lymphocyte recovery rates with significantly less platelet contamination than could be achieved using standard manual methods. In addition, automated processing was performed in a closed-system disposable apheresis kit, reducing the potential for bacterial contamination of the cell product. Culture of the lymphocytes in flexible plastic bags resulted in reductions in incubator space and culture media, and improved ease of handling of the cells. Lastly, automated harvest of cells out of culture led to increases in cell recovery without any damage to cell viability. Use of standard blood bank and apheresis technology in the collection, processing, culture, and harvest of LAK cells can significantly increase the efficiency and reduce the complexity of LAK generation and safeguard the sterility of the final product. Adoption of these techniques by smaller medical centers should allow LAK therapy to find more widespread use in the treatment of cancer. This work was done in collaboration with the Surgery Branch, NCI, as an addendum to Protocol No. 86-C-50.

为了简化淋巴因子激活的杀伤细胞（LAK）的生产， 用于治疗用途的细胞，自动细胞分离器（CS-3000，Fenwal） 适用于：（1）处理大量全血，用于收集 从癌症患者中浓缩白细胞，（2）纯化淋巴细胞 通过自动Ficoll-Hypaque密度梯度沉降法进行分级，（3） 在血小板储存袋中培养淋巴细胞，和（4）收获， 从培养物中浓缩LAK细胞。近100个LAK细胞制剂 都是以这种方式处理的。自动密度梯度 沉降导致高淋巴细胞回收率， 血小板污染比使用标准手册所能达到的更少 方法.此外，在封闭系统中进行自动化处理 一次性单采试剂盒，减少细菌感染的可能性 细胞产物的污染。淋巴细胞在软组织中的培养 塑料袋导致培养箱空间和培养基的减少， 并且提高了电池的处理容易性。最后，自动收获 细胞的培养导致细胞恢复的增加，而没有任何损害 to cell细胞viability活力. 在采集过程中使用标准血库和单采技术， LAK细胞的处理、培养和收获可以显著增加 效率和降低LAK产生和保障的复杂性 最终产品的无菌性。采用这些技术， 医疗中心应该允许LAK疗法在 癌症的治疗。 这项工作是与NCI外科分支合作完成的， 第86-C-50号议定书增编。