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Optimizing Donor Cellular Dissociation and Subretinal Injection Parameters for Stem Cell-Based Treatments.
- Source :
-
Stem cells translational medicine [Stem Cells Transl Med] 2019 Aug; Vol. 8 (8), pp. 797-809. Date of Electronic Publication: 2019 Apr 19. - Publication Year :
- 2019
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Abstract
- Subretinal delivery of stem cell-derived retinal cells as a strategy to treat retinal degenerative blindness holds great promise. Currently, two clinical trials are underway in which human fetal retinal progenitor cells (RPCs) are being delivered to patients by intravitreal or subretinal injection to preserve or restore vision, respectively. With the advent of the induced pluripotent stem cell (iPSC), and in turn three-dimensional derivation of retinal tissue, it is now possible to generate autologous RPCs for cell replacement. The purpose of this study was to evaluate the effect of commonly used cell isolation and surgical manipulation strategies on donor cell viability. iPSC-RPCs were subjected to various conditions, including different dissociation and isolation methods, injection cannula sizes, and preinjection storage temperatures and times. The effects of commonly used surgical techniques on both host and donor cell viability were evaluated in Yucatan mini-pigs (n = 61 eyes). We found a significant increase in cell viability when papain was used for RPC isolation. In addition, a significant decrease in cell viability was detected when using the 41G cannula compared with 31G and at storage times of 4 hours compared with 30 minutes. Although 96.4% of all eyes demonstrated spontaneous retinal reattachment following injection, retinal pigment epithelium (RPE) abnormalities were seen more frequently in eyes receiving injections via a 31G cannula; interestingly, eyes that received cell suspensions were relatively protected against such RPE changes. These findings indicate that optimization of donor cell isolation and delivery parameters should be considered when developing a subretinal cell replacement strategy. Stem Cells Translational Medicine 2019;8:797&809.<br /> (© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.)
- Subjects :
- Animals
Cell Survival
Cells, Cultured
Humans
Induced Pluripotent Stem Cells cytology
Induced Pluripotent Stem Cells transplantation
Neural Stem Cells cytology
Papain pharmacology
Primary Cell Culture standards
Retina drug effects
Stem Cell Transplantation adverse effects
Swine
Swine, Miniature
Cellular Reprogramming Techniques methods
Neural Stem Cells transplantation
Primary Cell Culture methods
Retina cytology
Retinal Dystrophies therapy
Stem Cell Transplantation methods
Subjects
Details
- Language :
- English
- ISSN :
- 2157-6580
- Volume :
- 8
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Stem cells translational medicine
- Publication Type :
- Academic Journal
- Accession number :
- 31004408
- Full Text :
- https://doi.org/10.1002/sctm.18-0210