Fate Therapeutics, Inc., 3535 General Atomics Court, Suite 200, San Diego, CA 92121, USA.
Stem Cell Reports. 2014 Mar 6;2(3):366-81. doi: 10.1016/j.stemcr.2014.01.014. eCollection 2014 Mar 11.
Cell banking, disease modeling, and cell therapy applications have placed increasing demands on hiPSC technology. Specifically, the high-throughput derivation of footprint-free hiPSCs and their expansion in systems that allow scaled production remains technically challenging. Here, we describe a platform for the rapid, parallel generation, selection, and expansion of hiPSCs using small molecule pathway inhibitors in stage-specific media compositions. The platform supported efficient and expedited episomal reprogramming using just OCT4/SOX2/SV40LT combination (0.5%-4.0%, between days 12 and 16) in a completely feeder-free environment. The resulting hiPSCs are transgene-free, readily cultured, and expanded as single cells while maintaining a homogeneous and genomically stable pluripotent population. hiPSCs generated or maintained in the media compositions described exhibit properties associated with the ground state of pluripotency. The simplicity and robustness of the system allow for the high-throughput generation and rapid expansion of a uniform hiPSC product that is applicable to industrial and clinical-grade use.
细胞库、疾病建模和细胞治疗应用对 hiPSC 技术提出了越来越高的要求。具体来说，在允许规模化生产的系统中，无足迹 hiPSC 的高通量衍生及其扩增在技术上仍然具有挑战性。在这里，我们描述了一个使用小分子通路抑制剂在特定阶段的培养基成分中快速、平行生成、选择和扩增 hiPSC 的平台。该平台支持高效和快速的瞬时重编程，仅使用 OCT4/SOX2/SV40LT 组合（12-16 天之间，0.5%-4.0%）在完全无饲养层的环境中进行。所得的 hiPSC 是无转基因的，容易培养和扩增为单细胞，同时保持同质和基因组稳定的多能性群体。在所述培养基组成中生成或维持的 hiPSC 表现出与多能性基础状态相关的特性。该系统的简单性和稳健性允许高通量生成和快速扩增均匀的 hiPSC 产物，适用于工业和临床级用途。
