Over the past decade, induced pluripotent stem cells (iPSCs) have become a major focus of stem cell and developmental biology research, offering researchers a clinically relevant source of cells that are amenable to genetic engineering approaches. Though stem cells are promising for both research and commercial endeavors, iPSC-based assays require tedious protocols that include complex treatments, expensive reagents, and specialized equipment that limit their integration into academic curricula and cell biology research groups. Expanding on existing Kit-On-A-Lid-Assay (KOALA) technologies, we have developed a self-contained, injection molded, pipette-less iPSC culture and differentiation platform that significantly reduces associated costs and labor of stem cell maintenance and differentiation. The KOALA kit offers users the full range of iPSC culture necessities, including cell cryopreservation, media exchanges, differentiation, endpoint analysis, and a new capability, cell passaging. Using the KOALA kit, we were able to culture ~20,000 iPSCs per microchannel for at least 7 days, while maintaining stable expression of stemness markers (SSEA4 and Oct4) and normal iPSC phenotype. We also adapted protocols for differentiating iPSCs into neuroepithelial cells, cardiomyocytes, and definitive endodermal cells, a cell type from each germ layer of human development.

在过去的十年中，诱导多能干细胞（iPSC）已成为干细胞和发育生物学研究的主要焦点，为研究人员提供了适合基因工程方法的临床相关细胞来源。尽管干细胞在研究和商业方面都有前景，但基于 iPSC 的检测需要繁琐的实验方案，包括复杂的处理、昂贵的试剂和专门的设备，限制了它们融入学术课程和细胞生物学研究小组。在现有的 Kit-On-A-Lid-Assay (KOALA) 技术的基础上，我们开发了一种独立、注塑、无移液器的 iPSC 培养和分化平台，可显着降低干细胞维护和分化的相关成本和劳动力。 KOALA 试剂盒为用户提供全方位的 iPSC 培养必需品，包括细胞冷冻保存、培养基交换、分化、终点分析和新功能（细胞传代）。使用 KOALA 试剂盒，我们能够在每个微通道中培养约 20,000 个 iPSC 至少 7 天，同时保持干性标记（SSEA4 和 Oct4）的稳定表达和正常的 iPSC 表型。我们还调整了将 iPSC 分化为神经上皮细胞、心肌细胞和定形内胚层细胞（人类发育的每个胚层的细胞类型）的方案。