Journal article
Propensity of Patient-Derived iPSCs for Retinal Differentiation: Implications for Autologous Cell Replacement
Stem cells translational medicine, Vol.12(6), pp.365-378
06/2023
DOI: 10.1093/stcltm/szad028
PMCID: PMC10267581
PMID: 37221451
Abstract
Prior to use, newly generated induced pluripotent stem cells (iPSC) should be thoroughly validated. While excellent validation and release testing assays designed to evaluate potency, genetic integrity, and sterility exist, they do not have the ability to predict cell type-specific differentiation capacity. Selection of iPSC lines that have limited capacity to produce high-quality transplantable cells, places significant strain on valuable clinical manufacturing resources. The purpose of this study was to determine the degree and root cause of variability in retinal differentiation capacity between cGMP-derived patient iPSC lines. In turn, our goal was to develop a release testing assay that could be used to augment the widely used ScoreCard panel. IPSCs were generated from 15 patients (14-76 years old), differentiated into retinal organoids, and scored based on their retinal differentiation capacity. Despite significant differences in retinal differentiation propensity, RNA-sequencing revealed remarkable similarity between patient-derived iPSC lines prior to differentiation. At 7 days of differentiation, significant differences in gene expression could be detected. Ingenuity pathway analysis revealed perturbations in pathways associated with pluripotency and early cell fate commitment. For example, good and poor producers had noticeably different expressions of OCT4 and SOX2 effector genes. QPCR assays targeting genes identified via RNA sequencing were developed and validated in a masked fashion using iPSCs from 8 independent patients. A subset of 14 genes, which include the retinal cell fate markers RAX, LHX2, VSX2, and SIX6 (all elevated in the good producers), were found to be predictive of retinal differentiation propensity.
Details
- Title: Subtitle
- Propensity of Patient-Derived iPSCs for Retinal Differentiation: Implications for Autologous Cell Replacement
- Creators
- Jessica A Cooke - University of IowaAndrew P Voigt - University of IowaMichael A Collingwood - Integrated DNA Technologies (United States)Nicholas E Stone - University of IowaS Scott Whitmore - University of IowaAdam P DeLuca - University of IowaErin R Burnight - University of IowaKristin R Anfinson - University of IowaChristopher A Vakulskas - Integrated DNA Technologies (United States)Austin J Reutzel - University of IowaHeather T Daggett - University of IowaJeaneen L Andorf - University of IowaEdwin M Stone - University of IowaRobert F Mullins - University of IowaBudd A Tucker - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Stem cells translational medicine, Vol.12(6), pp.365-378
- DOI
- 10.1093/stcltm/szad028
- PMID
- 37221451
- PMCID
- PMC10267581
- NLM abbreviation
- Stem Cells Transl Med
- ISSN
- 2157-6564
- eISSN
- 2157-6580
- Grant note
- R01 EY033331 / NEI NIH HHS
- Language
- English
- Electronic publication date
- 05/23/2023
- Date published
- 06/2023
- Academic Unit
- The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Center for Bioinformatics and Computational Biology; Ophthalmology and Visual Sciences
- Record Identifier
- 9984419359802771
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