Investigating the feasibility of scale up and automation of human induced pluripotent stem cells cultured in aggregates in feeder free conditions.
The transfer of a laboratory process into a manufacturing facility is one of the most critical steps required for the large scale production of cell-based therapy products. This study describes the first published protocol for scalable automated expansion of human induced pluripotent stem cell lines growing in aggregates in feeder-free and chemically defined medium. Cells were successfully transferred between different sites representative of research and manufacturing settings; and passaged manually and using the CompacT SelecT automation platform. Modified protocols were developed for the automated system and the management of cells aggregates (clumps) was identified as the critical step. Cellular morphology, pluripotency gene expression and differentiation into the three germ layers have been used compare the outcomes of manual and automated processes.
Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, University of Cambridge, UK; Centre for Biological Engineering, Loughborough University, UK. Electronic address: facs2@cam.ac.uk.
Filipa A C
FA
Soares
Centre for Biological Engineering, Loughborough University, UK.
Amit
A
Chandra
Centre for Biological Engineering, Loughborough University, UK.
Robert J
RJ
Thomas
Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, University of Cambridge, UK.
Roger A
RA
Pedersen
Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Anne McLaren Laboratory for Regenerative Medicine and Department of Surgery, University of Cambridge, UK; Wellcome Trust Sanger Institute, Hinxton, UK.
Ludovic
L
Vallier
Centre for Biological Engineering, Loughborough University, UK.
David J
DJ
Williams
D015415:Biomarkers / Q000032:analysis*; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002454:Cell Differentiation; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D004867:Equipment Design; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008856:Microscopy, Fluorescence; D012371:Robotics
D015415:Biomarkers; D003470:Culture Media
Automation; Feeder free; Induced pluripotent stem cells; Manufacturing; Scale up