STTR Phase I: Polymer electrosonic actuation microarray: Low-cost tool for transfection of biological cells

这个小企业技术转让（STTR）第一阶段项目将证明用于生物细胞机电转染的一次性聚合物微阵列的技术和商业可行性。需要替代基因转移解决方案来获得生物制药，基因治疗和干细胞研究的潜力。虽然用于基因转移的物理方法（例如，电穿孔）已经证明了在难以转染的原代细胞和干细胞中改善的（相对于常规化学介导的技术）治疗结果，但治疗的控制和均匀性仍然不足。此外，基于比色皿的电穿孔系统的性能的增量改进仅通过系统复杂性和成本的相应增加来实现。已经发现，细胞通过细胞大小的孔口一个接一个地排出，通过对每个单独的细胞施加相同且仔细控制的机电环境，促进细胞膜穿孔和DNA递送到细胞中。不幸的是，相对高的材料成本和对可实现的处理条件的限制（由于对使用标准硅微机械加工技术制造的微阵列的几何形状的限制）抑制了现有硅基微阵列的实际实施。STTR第一阶段项目中的聚合物微阵列的创新设计和优化将产生一个低成本的系统，该系统能够产生所需的机械应力场，以改善难以转染细胞的治疗效果。该项目更广泛的影响/商业潜力是使难以转染细胞的有效和经济的转染成为可能。在生命科学中，将原代细胞和干细胞用于各种研究和治疗应用。将基因、药物分子、成像剂、肽、抗体和酶有效递送到活细胞中对于从通过引入DNA治疗人类疾病到通过单分子成像研究基本细胞功能的应用范围是至关重要的;然而，细胞内递送和转染仍然是困难的任务。虽然在使用化学或物理方法来检测实验室建立和维持的（“容易”）细胞系的基础研究应用中，效率大于90%是常见的，但对于“困难”细胞，效率可能下降到10%或更低。物理方法的改进（例如，电穿孔）已经实现了渐进的性能改进;然而，目前市场上没有一个系统满足最终用户对效率、可行性、功能性和成本的所有要求。新的转染方法是STTR第一阶段项目的主题，它有望通过同时创新使用多种基因转移技术来提高治疗效果，同时通过为困难细胞提供具有成本效益的转染解决方案来更好地满足最终用户的需求。

This Small Business Technology Transfer (STTR) Phase I project will prove the technical and commercial feasibility of a single-use polymer microarray for electromechanical transfection of biological cells. Alternative gene transfer solutions are needed to access the potential held in biopharmaceuticals, gene therapy, and stem cell research. Although physical methods for gene transfer (e.g., electroporation) have demonstrated improved (over conventional chemical-mediated techniques) treatment outcomes in difficult-to-transfect primary and stem cells, control and uniformity of treatment remain inadequate. Further, incremental improvements in performance of cuvette-based electroporation systems have only been realized through corresponding increases in system complexity and cost. One-by-one ejection of cells through cell-sized orifices has been found to promote cell membrane poration and DNA delivery into cells by imposing an identical and carefully controlled electromechanical environment on each individual cell. Unfortunately, a relatively high material cost and limitations on achievable treatment conditions (due to constraints on the geometry of microarrays manufactured using standard silicon micromachining techniques), inhibit practical implementation of existing silicon-based microarrays. The innovative design and optimization of the polymer microarray under this STTR Phase I project will yield a low-cost system capable of generating the mechanical stress field needed to achieve improved treatment outcomes in difficult-to-transfect cells.The broader impact/commercial potential of this project is to enable effective and economical transfection of difficult-to-transfect primary and stem cells used for a variety of research and therapeutic applications in the Life Sciences. Effective delivery of genes, drug molecules, imaging agents, peptides, antibodies, and enzymes into living cells is critical to applications ranging from the treatment of human disease through introduction of DNA to the investigation of basic cellular function through single molecule imaging; yet, intracellular delivery and transfection remain difficult tasks. While efficiencies of greater than 90% are common in basic research applications that use chemical or physical methods to transfect laboratory established and maintained ("easy") cell lines, efficiency can drop to 10% or lower for "difficult" cells. Refinements of physical methods (e.g., electroporation) have achieved incremental performance improvements; however, no system currently on the market meets all end-user requirements for efficiency, viability, functionality, and cost. The novel approach to transfection, which is the subject of this STTR Phase I project, promises to improve treatment efficacy through innovative use of multiple gene transfer techniques simultaneously, while better addressing end user needs by providing a cost-effective transfection solution for difficult cells.