POLYETHYLENE GLYCOL AND ELECTRIC INDUCED MEMBRANE FUSION

聚乙二醇和电诱导膜融合

• 批准号: 2175966
• 负责人: SEK-WEN HUI
• 金额: $ 19.31万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2175966
• 关键词: cell fusion; electric field; electroporation; ethylene glycols; human tissue; hybridomas; laboratory mouse; laboratory rabbit; lipids; liposomes; membrane fusion; vesicle /vacuole

Our goal is to develop novel methods of artificially induced cell fusion to meet the growing needs of biomedicine and biotechnology. The molecular mechanisms of polyethylene glycol and electric field induced membrane fusion have been better understood, owing to our efforts and those of others in the past. This knowledge is to be used as basis to significantly improve the efficiency, versatility and controllability of new fusion methods. To improve fusion efficiency, cell contacts will be manipulated in centrifugation pellets, in overlaying monolayers, and in PEG-induced aggregates. The timing and degree of colloidal osmotic swelling after the pulse application, and the number of pulses will be controlled precisely to maximize fusion and viability, based on known membrane breakdown and cell swelling characteristics. By reducing cell lysis with polymers, and increasing contacts between heterogeneous fusion partners, we aim to induce efficient fusions between cells of different sizes, between suspending, adherent and tissue cells, as well as between liposomes, membrane vesicles and cells. Cationic lipids and PEG-conjugated lipids will be utilized to fine-tune the cell-liposome adhesion. A novel two phase polymer method will be used to enhance cell loading by electroporation. Highly efficient and controllable fusion methods will contribute to human hybridoma formation as well as in vitro gene transfer to human cell targets where cell sources are limited. Effective loading of targeted cells, or transferring membrane components to targeted cell surfaces by new fusion methods will be valuable in research and medical applications.

我们的目标是开发人工诱导细胞融合的新方法 以满足生物医学和生物技术日益增长的需求。 分子 聚乙二醇和电场诱导膜的机理 由于我们和其他组织的努力，融合已经得到了更好的理解 其他人过去。 这些知识将作为基础 显着提高效率、通用性和可控性 新的融合方法。 为了提高融合效率，细胞接触将被操纵 离心沉淀、重叠单层以及 PEG 诱导的 聚合体。 胶体渗透膨胀后的时间和程度 脉冲施加，脉冲数量精确控制 根据已知的膜破裂和 细胞肿胀特性。 通过用聚合物减少细胞裂解，以及 增加异质融合伙伴之间的联系，我们的目标是 诱导不同大小的细胞之间的有效融合 悬浮细胞、粘附细胞和组织细胞，以及脂质体之间， 膜囊泡和细胞。 阳离子脂质和 PEG 共轭脂质 将用于微调细胞-脂质体粘附。 小说二 相聚合物方法将用于增强细胞负载 电穿孔。 高效可控的融合方法将有助于人类 杂交瘤形成以及体外基因转移至人类细胞 细胞来源有限的目标。 有效装载目标 细胞，或通过以下方式将膜成分转移到目标细胞表面 新的融合方法将在研究和医学应用中有价值。