Closed, Aseptic Production Process for Engineered Living Human Skin Tissue

描述（由申请人提供）：组织工程（TE）有望对治疗数百万美国人的危及生命的疾病的治疗选择范围产生巨大影响。然而，迄今为止，还没有任何市售的TE产品被证明是商业上成功的。导致这种盈利能力缺乏的主要因素之一是所有步骤都需要无菌处理。与传统的制药和生物技术产品不同，工程组织和基于细胞的疗法必须在整个生产过程中进行无菌处理。因此，这些疗法具有极高的设施和生产成本。在美国医疗保健系统的患者可以使用基于细胞的疗法之前，需要新的、创新的无菌处理和放大方法。Stratatech公司已获得FDA批准，开始使用StrataGraft（TM）进行I/II期临床试验，StrataGraft（TM）是一种基于Stratatech专利的NIKS（TM）人角质形成细胞系的人皮肤替代组织。此外，Stratatech正在开发一种基因增强的活体皮肤产品（GliffGraft（tm）），用于慢性、非愈合性糖尿病、静脉淤滞和褥疮皮肤伤口市场。该快速通道提案旨在为这些创新产品开发一种封闭的无菌生产工艺，这将大大降低生产成本并提高患者安全性。在第一阶段，我们将解决封闭的，无菌的角质细胞培养和规模扩大的问题，以及PHS和FDA对异种移植的关注。我们将确定以无菌袋形式冷冻保存NIKS（tm）角质形成细胞的参数。然后，我们将在封闭的细胞工厂中解冻并繁殖这些保存的NIKS（tm）细胞，而无需小鼠饲养细胞的帮助。最后，我们将扩大NIKS（tm）培养的方法，以实现足够规模的收获和传代，用于全规模商业生产。在第二阶段，我们将通过开发在封闭的组织生长室中生产StrataGraft（TM）皮肤组织的方法来扩展这些结果。我们将构建原型室，并测试它们在批量和灌注模式下生产皮肤组织的能力，并进行在线过程监测。然后，我们将设计一个优化的第二代组织生长室，它也将作为最终的储存容器。相关性：该提案将开发创新的封闭式无菌处理方法，用于生产Stratatech Corporation正在开发的工程化活体人类皮肤替代产品StrataGraft（tm）和StrataGraft（tm）皮肤组织。通过以封闭、无菌的方式生产这些产品，而不需要小鼠饲养细胞，我们将把这些突破性的疗法带给数百万患有慢性、不可治愈的糖尿病、静脉淤滞和压力性溃疡的美国人。

DESCRIPTION (provided by applicant): Tissue engineering (TE) is poised to make an enormous impact on the range of therapeutic options for treatment of life-threatening conditions that afflict millions of Americans. To date, however, no marketed TE product has proven to be commercially successful. One of the primary factors contributing to this lack of profitability is the need for aseptic processing at all steps. Unlike traditional pharmaceutical and biotechnology products, engineered tissues and cell-based therapies must be processed aseptically throughout their manufacture. For this reason, these therapies come with extremely high facility and production costs. New, innovative approaches for aseptic processing and scale-up are required before cell- based therapies can become accessible to patients in the US health care system. Stratatech Corporation has approval from the FDA to begin Phase I/I I clinical trials with StrataGraft(tm), a human skin substitute tissue based on Stratatech's patented NIKS(tm) human keratinocyte cell line. In addition, Stratatech is developing a pipeline of genetically enhanced living skin products (ExpressGraft(tm)) for use in the chronic, non-healing diabetic, venous stasis, and decubitus skin wound markets. This Fast Track proposal is designed to develop a closed, aseptic manufacturing process for these innovative products which will greatly reduce production costs and increase patient safety. In Phase I we will address the issue of closed, aseptic keratinocyte culture and scale-up as well as the PHS and FDA concerns regarding xenotransplantation. We will determine parameters for cryopreservation of NIKS(tm) keratinocytes in sterile bag format. We will then thaw and propagate these preserved NIKS(tm) cells in closed cell factories without the aid of murine feeder cells. Finally, we will scale-up methods for NIKS(tm) culture to enable harvest and passage at a sufficient scale for full scale commercial manufacture. In Phase II we will extend these results by developing methods for production of StrataGraft(tm) skin tissue in closed tissue growth chambers. We will construct prototype chambers and test their ability to produce skin tissue in batch and perfusion modes with on-line process monitoring. Then, we will design an optimized second-generation tissue growth chamber which would also function as the final storage container. RELEVANCE: This proposal will develop innovative closed, aseptic processing methods for the manufacture of engineered living human skin substitute products, StrataGraft(tm) and ExpressGraft(tm) skin tissue, in development at Stratatech Corporation. By manufacturing these products in a closed, aseptic fashion without the need for mouse feeder cells, we will bring these breakthrough therapies to millions of Americans afflicted with chronic, non-healing diabetic, venous stasis, and pressure ulcers.