A Full Good Manufacturing Practice–Compliant Protocol for Corneal Stromal Stem Cell Cultivation
Mithun Santra,
Yen Hsu,
Shaheen Khadem,
Sydney Radencic,
Martha Funderburgh,
Onkar Sawant,
Deepinder Dhaliwal,
Vishal Jhanji,
Gary Yam
Affiliations
Mithun Santra
Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA
Yen Hsu
Immunologic Monitoring and Cellular Products Laboratory, Hillman Cancer Centre, University of Pittsburgh, Pittsburgh, PA, USACenter for Cell and Gene Therapy, Gift of Life Marrow Registry, Boca Raton, FL, USA
Shaheen Khadem
Immunologic Monitoring and Cellular Products Laboratory, Hillman Cancer Centre, University of Pittsburgh, Pittsburgh, PA, USA
Sydney Radencic
Immunologic Monitoring and Cellular Products Laboratory, Hillman Cancer Centre, University of Pittsburgh, Pittsburgh, PA, USA
Martha Funderburgh
Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA
Onkar Sawant
Center for Vision and Eye Banking Research, Eversight, Cleveland, OH, USA
Deepinder Dhaliwal
Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA
Vishal Jhanji
Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA
Gary Yam
Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USAMcGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA
Corneal scarring, a significant cause of global blindness, results from various insults, including trauma, infections, and genetic disorders. The conventional treatment to replace scarred corneal tissues includes partial or full-thickness corneal transplantation using healthy donor corneas. However, only 1 in 70 individuals with treatable corneal scarring can undergo surgery, due to the limited supply of transplantable donor tissue. Our research focuses on cell-based strategies, specifically ex vivo–expanded corneal stromal stem cells (CSSCs), to address corneal scarring. Preclinical studies have demonstrated the efficacy of CSSC treatment in reducing corneal inflammation and fibrosis, inhibiting scar formation, and regenerating native stromal tissue. Mechanisms include CSSC differentiation into stromal keratocytes and the expression of regenerative cytokines. Here, we present a good manufacturing practice (GMP)-compliant protocol to isolate and expand human CSSCs. This method paves the way to produce clinical-grade CSSCs for transplantation and clinical trials.
