Journal article
Automating iPSC generation to enable autologous photoreceptor cell replacement therapy
Journal of translational medicine, Vol.21(1), 161
02/28/2023
DOI: 10.1186/s12967-023-03966-2
PMCID: PMC9976478
PMID: 36855199
Abstract
Inherited retinal degeneration is a leading cause of incurable vision loss in the developed world. While autologous iPSC mediated photoreceptor cell replacement is theoretically possible, the lack of commercially available technologies designed to enable high throughput parallel production of patient specific therapeutics has hindered clinical translation.
In this study, we describe the use of the Cell X precision robotic cell culture platform to enable parallel production of clinical grade patient specific iPSCs. The Cell X is housed within an ISO Class 5 cGMP compliant closed aseptic isolator (Biospherix XVivo X2), where all procedures from fibroblast culture to iPSC generation, clonal expansion and retinal differentiation were performed.
Patient iPSCs generated using the Cell X platform were determined to be pluripotent via score card analysis and genetically stable via karyotyping. As determined via immunostaining and confocal microscopy, iPSCs generated using the Cell X platform gave rise to retinal organoids that were indistinguishable from organoids derived from manually generated iPSCs. In addition, at 120 days post-differentiation, single-cell RNA sequencing analysis revealed that cells generated using the Cell X platform were comparable to those generated under manual conditions in a separate laboratory.
We have successfully developed a robotic iPSC generation platform and standard operating procedures for production of high-quality photoreceptor precursor cells that are compatible with current good manufacturing practices. This system will enable clinical grade production of iPSCs for autologous retinal cell replacement.
Details
- Title: Subtitle
- Automating iPSC generation to enable autologous photoreceptor cell replacement therapy
- Creators
- Laura R Bohrer - University of IowaNicholas E Stone - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, USANathaniel K Mullin - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, USAAndrew P Voigt - University of Iowa, Ophthalmology and Visual SciencesKristin R Anfinson - University of IowaJessica L Fick - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, USAViviane Luangphakdy - Cell X Technologies Inc, Cleveland, OH, USABradley Hittle - The Ohio State UniversityKimerly Powell - The Ohio State UniversityGeorge F Muschler - Cleveland ClinicRobert F Mullins - University of IowaEdwin M Stone - University of IowaBudd A Tucker - Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, USA. budd-tucker@uiowa.edu
- Resource Type
- Journal article
- Publication Details
- Journal of translational medicine, Vol.21(1), 161
- DOI
- 10.1186/s12967-023-03966-2
- PMID
- 36855199
- PMCID
- PMC9976478
- NLM abbreviation
- J Transl Med
- ISSN
- 1479-5876
- eISSN
- 1479-5876
- Grant note
- RO1 EY033331 / NIH HHS
- Language
- English
- Date published
- 02/28/2023
- Academic Unit
- The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Ophthalmology and Visual Sciences
- Record Identifier
- 9984375455202771
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