TY - JOUR
T1 - Automating iPSC generation to enable autologous photoreceptor cell replacement therapy
AU - Bohrer, Laura R.
AU - Stone, Nicholas E.
AU - Mullin, Nathaniel K.
AU - Voigt, Andrew P.
AU - Anfinson, Kristin R.
AU - Fick, Jessica L.
AU - Luangphakdy, Viviane
AU - Hittle, Bradley
AU - Powell, Kimerly
AU - Muschler, George F.
AU - Mullins, Robert F.
AU - Stone, Edwin M.
AU - Tucker, Budd A.
N1 - Funding Information:
RNA-seq data presented herein were generated by the Genomics Division of the Iowa Institute of Human Genetics which is supported, in part, by the University of Iowa Carver College of Medicine.
Funding Information:
This research was funded by the National Institutes of Health R01 EY033331.
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/12
Y1 - 2023/12
N2 - Background: 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. Methods: 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. Results: 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. Conclusion: 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.
AB - Background: 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. Methods: 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. Results: 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. Conclusion: 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.
KW - Automation
KW - Cell manufacturing
KW - Cell therapy
KW - Induced pluripotent stem cells
KW - RNA sequencing
KW - Retinal differentiation
KW - Robotic cell culture
UR - http://www.scopus.com/inward/record.url?scp=85149153165&partnerID=8YFLogxK
U2 - 10.1186/s12967-023-03966-2
DO - 10.1186/s12967-023-03966-2
M3 - Article
C2 - 36855199
AN - SCOPUS:85149153165
SN - 1479-5876
VL - 21
JO - Journal of Translational Medicine
JF - Journal of Translational Medicine
IS - 1
M1 - 161
ER -