Abstract
Industrialization of stem-cell based therapies requires innovative solutions to close the gap between research and commercialization. Scalable cell production platforms are needed to reliably deliver the cell quantities needed during the various stages of development and commercial supply. Human pluripotent stem cells (hPSCs) are a key source material for generating therapeutic cell types. We have developed a closed, automated and scalable stirred tank bioreactor platform, capable of sustaining high fold expansion of hPSCs. Such a platform could facilitate the in-process monitoring and integration of online monitoring systems, leading to significantly reduced labor requirements and contamination risk. hPSCs are expanded in a controlled bioreactor using perfused xeno-free media. Cell harvest and concentration are performed in closed steps. The hPSCs can be cryopreserved to generate a bank of cells, or further processed as needed. Cryopreserved cells can be thawed into a two-dimensional (2D) tissue culture platform or a three-dimensional (3D) bioreactor to initiate a new expansion phase, or be differentiated to the clinically relevant cell type. The expanded hPSCs express hPSC-specific markers, have a normal karyotype and the ability to differentiate to the cells of the three germ layers. This end-to-end platform allows a large scale expansion of high quality hPSCs that can support the required cell demand for various clinical indications.
Highlights
Successful generation of human-induced pluripotent stem cells by somatic cell reprogramming has opened new avenues in regenerative medicine, disease modeling and drug development [1,2]
The results indicate that L7TM TFO2 media supports MC-based expansion of Human pluripotent stem cells (hPSCs) in suspension
Cells that were seeded with uncoated MCs failed to show growth and expansion (Figure 2I–J). These results demonstrate that coating the MCs with L7TM hPSC Matrix is required for cell expansion
Summary
Successful generation of human-induced pluripotent stem cells (hiPSCs) by somatic cell reprogramming has opened new avenues in regenerative medicine, disease modeling and drug development [1,2]. Capable of self-renewal and pluripotency, hiPSCs derived from patients of both normal and aberrant phenotypes provide a theoretically limitless supply of clinically relevant iPSC-derived cells without ethical limitations and immune-rejection [3,4]. Given the heart’s limited-to-no regenerative capacity, new cardiomyocytes can be derived from hiPSCs by modulating developmental cues critical in embryonic development in vivo [5,6]. Essential to the successful differentiation of iPSCs to a specific cell lineage, includes careful consideration of the microenvironment and method with which iPSCs are maintained.
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