Automated, scalable culture of human embryonic stem cells in feeder‐free conditions

Abstract

Large-scale manufacture of human embryonic stem cells (hESCs) is prerequisite to their widespread use in biomedical applications. However, current hESC culture strategies are labor-intensive and employ highly variable processes, presenting challenges for scaled production and commercial development. Here we demonstrate that passaging of the hESC lines, HUES7, and NOTT1, with trypsin in feeder-free conditions, is compatible with complete automation on the CompacT SelecT, a commercially available and industrially relevant robotic platform. Pluripotency was successfully retained, as evidenced by consistent proliferation during serial passage, expression of stem cell markers (OCT4, NANOG, TRA1-81, and SSEA-4), stable karyotype, and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Automation of hESC culture will expedite cell-use in clinical, scientific, and industrial applications.

Multiparameter flow cytometry for the characterization of human embryonic stem cells

“…HUES7 cells were cultured feeder free on Matrigel TM basement matrix ((BD Biosciences) Thomas et al 2009). Monolayer cultures were maintained in 5% CO 2 37°C incubators between passages 24 and 36.…”

Precision manufacturing for clinical-quality regenerative medicines

“…Successful automated production protocols were developed for the expansion of cell numbers for human mesenchymal stem cells [16] and human embryonic stem cells (hESCs) [17], as these represent the cell stock source for a significant proportion of cell therapies under development. Further work with collaborators also developed automated production protocols for niche proprietary cell lines and Figure 3.…”

Concise Review: Epidermal Grafting: The Case for Pluripotent Stem Cells

“…Several groups have attempted to integrate automation into culture of pluripotent stem cells, using bioreactors to grow the cells [19], mechanical passaging, automated cell seeding, etc. Finally, the full automation of cell culture over up to 10 passages led to the production of three billion cells on a CompacT SelecT platform [20], thus providing definitive demonstration for the scalability of the process. The authors of that study have calculated that, running continuously, one of those automated platforms would provide a manufacturing capacity of two billion cells every 2 days.…”