An Integrated Miniature Bioprocessing for Personalized Human Induced Pluripotent Stem Cell Expansion and Differentiation into Neural Stem Cells

Abstract

Human induced pluripotent stem cells (iPSCs) are ideal cell sources for personalized cell therapies since they can be expanded to generate large numbers of cells and differentiated into presumably all the cell types of the human body in vitro. In addition, patient specific iPSC-derived cells induce minimal or no immune response in vivo. However, with current cell culture technologies and bioprocessing, the cost for biomanufacturing clinical-grade patient specific iPSCs and their derivatives are very high and not affordable for majority of patients. In this paper, we explored the use of closed and miniature cell culture device for biomanufacturing patient specific neural stem cells (NSCs) from iPSCs. We demonstrated that, with the assist of a thermoreversible hydrogel scaffold, the bioprocessing including iPSC expansion, iPSC differentiation into NSCs, the subsequent depletion of undifferentiated iPSCs from the NSCs, and concentrating and transporting the purified NSCs to the surgery room, could be integrated and completed within two closed 15 ml conical tubes.

Scalable Generation of Mesenchymal Stem Cells from Human Embryonic Stem Cells in 3D

“…As alternative sources, hESC and iPSC can generate MSC theoretically for an unlimited time. Recently, substantial progress has been achieved to directly differentiate hESC spheroids into 3D structures such as cardiomyocytes 21 , neural progenitors and hepatocytes 28 , 29 . Without the need for culture splits (which are associated with repeat cell detachment, dissociation, and reattachment), these 3D direct differentiation methods have fewer cell loss, save space, time, supplies, and labor, thus having great advantages over 2D differentiation systems and favoring large-scale production of the therapeutic cells in a bioreactor.…”

A Scalable and Efficient Bioprocess for Manufacturing Human Pluripotent Stem Cell-Derived Endothelial Cells

“…The hydrogel scaffold protects cells from hydrodynamic stresses in the culture vessel and prevents cells from excessive agglomeration, leading to high culture efficiency. For instance, the hydrogel scaffold enables long-term, serial expansion of hPSCs with a high cell viability (e.g., >90%, Figures 1 D, S2 C, and S2F), growth rate (e.g., 20-fold/5days, Figure 1 E), yield (e.g., 2.0 × 10 7 cells/mL, Figure 1 F), and purity (>99%, Figure 1 C, S2 B, and S2E), all of which offer considerable improvements over 3D suspension cultures ( Lei and Schaffer, 2013 , Li et al., 2016 , Lin et al., 2017 ). We hypothesize that hPSCs can also be differentiated into ECs in this culture system.…”

“…To address the challenge, we previously developed a scalable, efficient, and current Good Manufacturing Practice (cGMP)-compliant method for expanding hPSCs ( Lei and Schaffer, 2013 , Li et al., 2016 , Lin et al., 2017 ). The method, which was successfully repeated in this study ( Figures 1 and S2 ), uses a 3D thermoreversible hydrogel (Mebiol Gel) as the scaffold.…”

Advanced Materials to Enhance Central Nervous System Tissue Modeling and Cell Therapy

“…This method increased cell survival compared to standard 3D bioreactors with free-floating cells by both reducing hydrodynamic stresses and ensuring efficient nutrient transport via controlling the cluster size of expanding cells to less than 400 µm (radial diameter). [80] Together with higher cell yields and scalability compared to other culture platforms, this system also adds more control over stem cell differentiation by both alleviating agglomeration as well as allowing gentle cell retrieval by simply adding cold media to the hydrogel followed by partial dissociation of the cell clusters, removing the need for enzymatic cell harvesting and minimizing chemical and physical stresses.…”