Enhancing the Scalable Manufacturing of Human Mesenchymal Stromal Cells in Dynamic Culture Using Bioactive Heparin/Collagen Layer‐by‐Layer Coated Microcarriers
TL;DR
It is demonstrated that heparin/collagen Layer‐by‐Layer (LbL) coatings enhance hMSC functional performance and scalability of microcarrier‐based culture, especially at lower initial cell densities, representing a promising approach for scalable cell manufacturing with the potential for GMP adaptation.
Abstract
Expansion of human mesenchymal stromal cells (hMSCs) remains a bottleneck during manufacturing, where high cell yields must be achieved without compromising quality. Microcarriers provide elevated surface areas but often rely on synthetic materials. Here we use heparin/collagen Layer‐by‐Layer (LbL) coatings on microcarriers to enhance dynamic hMSC culture. HMSCs were cultured on LbL‐coated and untreated microcarriers in up to 500 mL flasks, and were assessed over time, up to 18 days across three seeding densities (2600–20,000 cells/cm2), while yield was assessed at the end of culture. Metabolic analysis suggests no induction of a generalized stress response, increased glycolytic activity, and a shift toward a more active secretory state for cells grown on coated surfaces, as measured by noninvasive assays (Luminex multiplex and Raman spectroscopy). At medium and high densities, the cost (18.6%) and yield (60% and 30%, respectively) increases (coated: untreated) were moderately favorable. However, at the lower density, the cost increase remained similar, despite a yield increase of 110%. Together, the findings of this study demonstrate that our LbL surface coatings enhance hMSC functional performance and scalability of microcarrier‐based culture, especially at lower initial cell densities, representing a promising approach for scalable cell manufacturing with the potential for GMP adaptation.