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Publications

Journal Article Epublish

PubMed: 42774489 DOI: 10.1016/j.mtbio.2026.103638

Robotic handling preserves iPSC-derived vascular smooth muscle cell differentiation.

Mater Today Bio 40:103638

Authors: Monique Bax, Jeya Ramalingam, Valentin Romanov, Muhammad A Alsherbiny, Keerat Junday, Jordan Thorpe, Emily Hurley, Melanie Lim, Ingrid Tarr, Stephanie Hesselson, Eleni Giannoulatou, Renjing Liu, Adam Hill, Siiri Iismaa, Robert M Graham

Affiliations

  • Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2010, Australia. School of Clinical Medicine, St Vincent's Healthcare Clinical Campus, Faculty of Medicine and Health, UNSW Sydney, Australia. Department of Medicine, University of Cambridge, Cambridge, United Kingdom. Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, Cambridge Biomedical Campus, University of Cambridge, Cambridge, United Kingdom.
  • Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2010, Australia.
  • Victor Chang Cardiac Research Institute, Darlinghurst, NSW, 2010, Australia. School of Clinical Medicine, St Vincent's Healthcare Clinical Campus, Faculty of Medicine and Health, UNSW Sydney, Australia.
  • School of Clinical Medicine, St Vincent's Healthcare Clinical Campus, Faculty of Medicine and Health, UNSW Sydney, Australia.
  • Department of Medicine, University of Cambridge, Cambridge, United Kingdom.
  • St Vincent's Hospital, Darlinghurst, NSW, 2010, Australia.

Abstract

To tackle the global cardiovascular disease epidemic, vascular cell culture requires scalable, efficient, and reproducible methods for differentiating cells. However, no studies have validated whether automation, essential for scale-up, preserves differentiation quality of cells. Here, we developed a differentiation protocol for induced pluripotent stem cell (iPSC)-derived vascular smooth muscle cells (iVSMCs) that is applicable to both manual, and automated robotic culture. iPSC lines generated from male and female donors across various ages were used to optimize the generation of iVSMCs. iVSMCs had consistent morphology and protein expression, which were largely similar to those of primary VSMCs, including expression of myosin heavy chain-11 (MYH11), α-smooth muscle actin (α-SMA), transgelin (TAGLN) and calponin (CNN1). Functionally, these iVSMCs responded to the vasoconstrictor carbachol. Coupling this protocol with an automated Hamilton liquid-handling robotics system allowed the generation of iVSMCs in large quantities, that were morphologically similar to manually differentiated iVSMCs. Comparative proteomic analysis confirmed that protein expression was substantially the same between automated and manual differentiation methods. Automation markedly reduced manual labor and facilitated increased production without sacrificing cell quality. This study demonstrates the feasibility of automating iVSMC differentiation, marking a significant step towards scalable VSMC manufacture for three-dimensional applications, and for the ever-growing demands of organoid and tissue engineering applications.

MeSH

Cells, Muscle Cells, Cell Differentiation, Muscles, Microscopy, Electron, Scanning Transmission, Muscle, Smooth, Muscle, Smooth, Vascular, Myocytes, Smooth Muscle