July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
3D hydrogels protect human retinal progenitor cells from stress exerted during transplantation
Author Affiliations & Notes
  • PIERRE COLOMBE
    Material Sciences and engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States
    Schepens Eye research Institute, Harvard Medical School, Boston, Massachusetts, United States
  • Deepti Singh
    Schepens Eye research Institute, Harvard Medical School, Boston, Massachusetts, United States
  • Myron Spector
    Material Sciences and engineering, Harvard Medical School, Boston, Massachusetts, United States
    Schepens Eye research Institute, Harvard Medical School, Boston, Massachusetts, United States
  • Michael J Young
    Schepens Eye research Institute, Harvard Medical School, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   PIERRE COLOMBE, None; Deepti Singh, None; Myron Spector, None; Michael Young, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 3328. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      PIERRE COLOMBE, Deepti Singh, Myron Spector, Michael J Young; 3D hydrogels protect human retinal progenitor cells from stress exerted during transplantation. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3328.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : The current challenges faced in cell therapy is a lack of approved polymeric cell carriers that could aid in delivery of cells and would enhance cell survival and degrade within an acceptable time frame without triggering an immune response from the host. This study is aimed at exploring the use of a hydrogel as a “protective envelope” for cells that experience high stress exerted during transplantation by doing, first, an invitro study to replicate the conditions of transplantation and, secondly, an invivo study to confirm those results.

Methods : Human retinal progenitor cells (hRPCs) were cultured in different condition on a 2D layer until reaching confluence (PBS with endothelial growth factor (EGF), cells encapsulated in gel with PBS and EGF). We have developed a gelatin-hydroxyphenyl propionic acid (GA-HPA) hydrogel with a gelation time of 20 seconds. To check viability, proliferation, apoptosis, stemness, retinal progenitor and cones markers immunohistostaining and flow cytometry were performed. Live and dead assay were used to study viability, Caspase 9 and Ki67 were used for apoptosis and proliferation. Finally flow cytometry was performed using MACSQuant to study stemness, retinal progenitor and cones markers. The Invivo study was performed on Long Evans rats by injecting in the subretinal space different conditions (hRPCs alone, cells encapsulated in gel with or without EGF).

Results : Live-dead assay shows cells encapsulated in gel have higher viability in comparison with non-gel cultures. Cells cultured with gel show higher viability and proliferation in comparison to the monolayer culture. Apoptosis quantification using CAS9 clearly indicated the protective properties of the gel. Flow cytometry assay shows cells encapsulated in gel express higher percentage of stemness and retinal progenitor markers. More viable cells were found in the 3D culture compared to saline-delivered controls. Furthermore, the in-vivo study shows higher integration and viability of cell injected in the 3D culture after 3 days.

Conclusions : The invitro and invivo results with gel have shown great promise and we intend to use it in the degenerative animal model to ehance the chances of cell survival and integration for restoring vision. This study demonstrates GA-HP injected in-situ crosslinking biodegradable hydrogel has the potential to be used as cell carrier for in-vivo delivery of stem and progenitor cells.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×