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Related Experiment Video

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Optimized Minimally Invasive Transscleral Subretinal Injection Technique in Mouse
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Optimizing Donor Cellular Dissociation and Subretinal Injection Parameters for Stem Cell-Based Treatments.

Brittni A Scruggs1, Chunhua Jiao1, Cathryn M Cranston1

  • 1University of Iowa Institute for Vision Research and the Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, Iowa, USA.

Stem Cells Translational Medicine
|April 21, 2019
PubMed
Summary

Optimizing cell isolation and delivery methods is crucial for subretinal stem cell therapy. Papain isolation and shorter storage times improve retinal progenitor cell (RPC) viability for treating degenerative blindness.

Keywords:
Induced pluripotent stem cellsInherited retinal dystrophyPig modelRetinal progenitor cellsStem cell transplantationSubretinal transplantation

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Area of Science:

  • Ophthalmology
  • Regenerative Medicine
  • Cell Biology

Background:

  • Subretinal delivery of stem cell-derived retinal cells shows promise for treating retinal degenerative blindness.
  • Current clinical trials utilize human fetal retinal progenitor cells (RPCs) for vision preservation or restoration.
  • Induced pluripotent stem cells (iPSCs) enable the generation of autologous RPCs for cell replacement therapy.

Purpose of the Study:

  • To evaluate the impact of cell isolation and surgical manipulation on donor cell viability for subretinal delivery.
  • To assess the effects of various dissociation methods, cannula sizes, and storage conditions on iPSC-RPCs.
  • To analyze the influence of surgical techniques on host and donor cell viability in a preclinical model.

Main Methods:

  • iPSC-RPCs were subjected to different dissociation and isolation techniques.
  • Viability was assessed after exposure to varying injection cannula sizes (31G vs. 41G).
  • Pre-injection storage temperatures and times (30 minutes vs. 4 hours) were evaluated, alongside surgical techniques in Yucatan mini-pigs.

Main Results:

  • Papain isolation significantly increased RPC viability compared to other methods.
  • A 41G cannula and 4-hour storage time significantly decreased cell viability.
  • While most eyes showed spontaneous retinal reattachment, 31G cannula injections correlated with more frequent retinal pigment epithelium (RPE) abnormalities, though cell suspensions offered protection.

Conclusions:

  • Optimization of donor cell isolation protocols, such as using papain, is essential for enhancing cell viability.
  • Delivery parameters, including cannula size and pre-injection storage duration, significantly impact cell survival and therapeutic outcomes.
  • Careful consideration of surgical manipulation techniques is critical for successful subretinal cell replacement strategies in treating retinal degenerative diseases.