Building 3D Culture Systems for Biomanufacturing Stem Cells at Low Cost

Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), have the capacities for indefinite in vitro expansion and differentiation into presumably all cell types in the human body. They therefore represent highly promising cell sources for numerous biomedical applications, such as cell replacement therapy, tissue and organ engineering, and pharmacology and toxicology screens. Many such applications require large numbers of cells of high quality and purity. For instance, ~10⁵ surviving dopaminergic neurons, ~10⁹ cardiomyocytes, or ~10⁹ β cells are required to treat a patient with Parkinson’s disease (PD), myocardial infarction (MI), or type I diabetes respectively. Analogously, ~10¹⁰ hepatocytes are needed for an artificial human liver, and ~10¹⁰cells may be required to screen a million compound library. Considering the large patient populations with degenerative diseases or organ failure (e.g. over 1 million with PD, 1-2.5 million with type I diabetes, ~8 million with MI, and 5.7 million with heart failure in the US alone), as well as the millions of chemical/peptide/nucleotide compounds that can be screened against many cell types, massive numbers of hPSCs are thus needed.

 It is becoming clear that the current 2D-based cell culture systems are incapable of producing sufficient cells with high quality and are becoming a bottleneck for these downstream applications. An attractive approach for scaling up cell production is to move the cell culture from 2D to 3D. cGMP compliant and scalable 3D culture systems that can biomanufacture high quality hPSCs and their derivatives at low cost are of great interest. In this presentation, we will introduce a simple, defined, scalable, cGMP compliant 3D culture system. With this system, hPSCs can be expanded 1000-fold over 11 days, resulting in extremely high volumetric yield. The high yield dramatically reduce the biomanufacturing cost. hPSCs can be cultured for long term (e.g. over 10 passages) without losing their pluripotency. 95% cells are viable and >95% cells are Oct+/Nanog+. hPSCs can be differentiated into various cell types in the system. The culture system can also be used to culture other tissue stem cells, such as cancer stem cells and chondrocytes.