A chemically-defined plastic scaffold for the xeno-free production of human pluripotent stem cells

Abstract

Clinical use of human pluripotent stem cells (hPSCs) is hampered by the technical limitations of their expansion. Here, we developed a chemically synthetic culture substrate for human pluripotent stem cell attachment and maintenance. The substrate comprises a hydrophobic polyvinyl butyral-based polymer (PVB) and a short peptide that enables easy and uniform coating of various types of cell culture ware. The coated ware exhibited thermotolerance, underwater stability and could be stored at room temperature. The substrate supported hPSC expansion in combination with most commercial culture media with an efficiency similar to that of commercial substrates. It supported not only the long-term expansion of examined iPS and ES cell lines with normal karyotypes during their undifferentiated state but also directed differentiation of three germ layers. This substrate resolves major concerns associated with currently used recombinant protein substrates and could be applied in large-scale automated manufacturing; it is suitable for affordable and stable production of clinical-grade hPSCs and hPSC-derived products.

Polymerized Laminin-521: A Feasible Substrate for Expanding Induced Pluripotent Stem Cells at a Low Protein Concentration

“…A different approach to cultivating hiPSCs under chemically defined conditions is the use of synthetic substrates such as polyvinyl alcohol (PVA), poly [poly (ethylene glycol) monomethacrylate] (PEGMA), and poly (N-isopropylacrylamide) (PNIPAM) [ 37 , 38 ]. Although synthetic polymers can be broadly applied on different surfaces and used to demonstrate that cells can be expanded and maintain their pluripotent characteristics, there are several technical limitations to this approach.…”

“…When Matrigel or LN511 is used, hPSCs predominantly adhere to the substrate via ITGA5 and ITGA6 and proliferate via β1 integrin [ 42 , 43 ]. In addition, when cells were cultivated on vitronectin substrate [ 38 ], ITGA3 expression correlated with cell attachment and migration, suggesting that the substrate affects these in vitro interactions. Given that polyLN521 assembles in homogeneous layers, we evaluated the effect of coating concentration on the cell expression of LAMA5 and integrins (i.e., ITGA3 and ITGA6) on day 2 and day 4 of expansion.…”

Cell Behavioral Dynamics as a Cue in Optimizing Culture Stabilization in the Bioprocessing of Pluripotent Stem Cells

“…The availability of synthetic polymer- and peptide-based matrices with tunable mechanical properties allows users to regulate an optimal strength of cell–substrate adhesion in a target cell-specific manner, thereby promoting an efficient generation of desired cells [ 35 , 79 , 85 , 86 , 87 , 88 ]. Stiffness represents a key mechanical property of coating material, critically dictating the subcellular allocation and activity of the integrin-mediated adhesion molecules, and further modifying the cell interactions with neighboring cells [ 89 ].…”

Progress and emerging techniques for biomaterial-based derivation of mesenchymal stem cells (MSCs) from pluripotent stem cells (PSCs)

“…A few of these scaffolds play an essential role in PSC proliferation and differentiation. A peptide polyvinyl butyral-based polymer scaffold synthesized by means of acrylic polymerization led to effective proliferation and differentiation of PSCs into cells of the mesodermal lineage, and therefore, it can potentially be useful for generating MSCs [ 198 ].…”