Closed, Aseptic Production Process for Engineered Living Human Skin Tissue

描述(由申请人提供)：组织工程学(TE)将对困扰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，到目前为止，还没有一种上市的TE产品被证明是商业上成功的。造成这种盈利能力不足的主要因素之一是需要在所有步骤进行无菌加工。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制造过程中进行无菌处理。出于这个原因，这些疗法的设施和生产成本都非常高。在美国医疗保健系统中的患者能够获得基于细胞的疗法之前，需要采用新的创新方法进行无菌处理和扩大规模。Stratech公司已经获得FDA的批准，可以开始StrataGraft(Tm)的I/I期临床试验，StrataGraft(Tm)是一种基于Stratech的专利NIKS(Tm)人角质形成细胞系的人类皮肤替代组织。此外，Stratech正在开发一系列基因增强型活性皮肤产品(ExpressGraft(Tm))，用于慢性、不可治愈的糖尿病、静脉淤滞和卧位症皮肤伤口市场。这项快速通道提案旨在为这些创新产品开发一种封闭的无菌制造工艺，这将极大地降低生产成本并提高患者的安全性。在第一阶段，我们将解决封闭的、无菌角质形成细胞培养和放大的问题，以及PHS和FDA对异种移植的担忧。我们将确定NIKS(TM)角质形成细胞在无菌袋中冷冻保存的参数。然后，我们将在封闭的细胞工厂中解冻和繁殖这些保存的NIKS(Tm)细胞，而不需要小鼠饲养细胞的帮助。最后，我们将扩大NIKS(Tm)培养方法的规模，使其能够以足够的规模收获和传代，以进行全面的商业生产。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。我们将建造样品室，并通过在线过程监测来测试它们批量和灌流模式生产皮肤组织的能力。然后，我们将设计一个优化的第二代组织生长室，它也将作为最终的存储容器。相关性：这项提案将为Stratech公司正在开发的工程活体人体皮肤替代品产品StrataGraft(Tm)和ExpressGraft(Tm)皮肤组织的生产开发创新的封闭、无菌加工方法。通过在不需要小鼠饲养细胞的情况下以封闭、无菌的方式制造这些产品，我们将把这些突破性疗法带给数百万患有慢性、无法愈合的糖尿病、静脉阻塞和压疮的美国人。

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.