Engineering a Human Organ for Dry Storage

设计用于干燥储存的人体器官

• 批准号: 6641852
• 负责人: Paul Barth Conrad
• 金额: $ 9.98万
• 依托单位: STRATATECH CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2003-12-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6641852
• 关键词: biotechnology; clinical research; expression cloning; keratinocyte; technology /technique development; temperature; tissue /cell culture; tissue /cell preparation; tissue /organ preservation; tissue engineering; transfection; transfection /expression vector; trehalose

DESCRIPTION (provided by applicant): Tissue engineering is a field with tremendous untapped potential, current revenues ($232 million in 2000, Genetic Engineering News 9/15/01) fall far short of the estimated market of $100 billion (Barron's, 5/15/00). This gap is in large part due to the significant barriers of short shelf life and high storage cost. Stratatech proposes to develop technology overcoming these hurdles using genetic manipulations of source cells coupled with recent advances in preservation solution engineering to create methods enabling room temperature storage of living cells and organs. Currently tissues are preserved by slow freezing during which they suffer numerous damaging effects. Rapid freezing or vitrification avoids these issues by preserving samples in an amorphous glass, but vitrification can give rise to lethal intracellular ice formation. Agents such as trehalose, a disaccharide of glucose, enhance the vitrification properties of preservation solutions; unfortunately, trehalose cannot penetrate cell membranes so novel methods must be used to deliver intracellular trehalose. Furthermore, until recently, vitrification of medically useful sample sized was not practical, but recent work has shown that careful engineering of the preservation solution can greatly enhance trehalose's ability to raise glass transition temperatures. Stratatech is developing a line of human keratinocytes for therapeutic applications called NIKS cells. NIKS cells are an ideal vehicle for this proposal in that they can be perpetually cultured in vitro and they can be used to manufacture a functional human organ. In addition they are pathogen-free, non-tumorigenic and they recapitulate the correct architecture of human skin. NIKS cells and skin tissue grown from them are capable of stable genetic modifications making it possible to optimize their survival in the dry state.

描述（由申请人提供）： 组织工程是一个具有巨大未开发潜力的领域，目前的收入（2000年为2.32亿美元，Genetic Engineering News 9/15/01）远远低于1000亿美元的估计市场（巴伦，5/15/00）。这一差距在很大程度上是由于保质期短和储存成本高的重大障碍。Stratatech提出开发克服这些障碍的技术，使用源细胞的遗传操作，加上保存溶液工程的最新进展，以创建能够在室温下储存活细胞和器官的方法。 目前，组织通过缓慢冷冻来保存，在此期间，它们遭受许多破坏性影响。快速冷冻或玻璃化通过将样品保存在无定形玻璃中来避免这些问题，但玻璃化可引起致命的细胞内冰形成。诸如海藻糖（葡萄糖的二糖）的试剂增强了保存溶液的玻璃化性质;不幸的是，海藻糖不能穿透细胞膜，因此必须使用新的方法来递送细胞内海藻糖。此外，直到最近，玻璃化的医学上有用的样品大小是不实际的，但最近的工作表明，精心设计的保存溶液可以大大提高海藻糖的能力，提高玻璃化转变温度。 Stratatech正在开发一种用于治疗应用的人类角质形成细胞，称为NIKS细胞。NIKS细胞是该提议的理想载体，因为它们可以在体外永久培养，并且它们可以用于制造功能性人体器官。此外，它们是无病原体的，非致瘤性的，并且它们概括了人类皮肤的正确结构。NIKS细胞和由其生长的皮肤组织能够进行稳定的遗传修饰，从而可以优化其在干燥状态下的存活。