Rapid bioprinting of conjunctival stem cell micro-constructs for subconjunctival ocular injection

Zheng Zhong; Xiaoqian Deng; Pengrui Wang; Claire Yu; Wisarut Kiratitanaporn; Xiaokang Wu; Jacob Schimelman; Min Tang; Alis Balayan; Emmie Yao; Jing Tian; Luwen Chen; Kang Zhang; Shaochen Chen

doi:10.1016/j.biomaterials.2020.120462

Rapid bioprinting of conjunctival stem cell micro-constructs for subconjunctival ocular injection

Biomaterials. 2021 Jan:267:120462. doi: 10.1016/j.biomaterials.2020.120462. Epub 2020 Oct 23.

Authors

Zheng Zhong¹, Xiaoqian Deng², Pengrui Wang³, Claire Yu¹, Wisarut Kiratitanaporn⁴, Xiaokang Wu⁵, Jacob Schimelman¹, Min Tang¹, Alis Balayan⁶, Emmie Yao¹, Jing Tian⁴, Luwen Chen¹, Kang Zhang⁷, Shaochen Chen⁸

Affiliations

¹ Department of NanoEngineering, University of California San Diego, La Jolla, CA, 92093, USA.
² Department of Human Genetics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, 90095, USA.
³ Materials Science and Engineering Program, University of California San Diego, La Jolla, CA, 92093, USA.
⁴ Department of Bioengineering, University of California San Diego, La Jolla, CA, 92093, USA.
⁵ Smidt Heart Institute, Cedars-Sinai Medical Center, Los Angeles, CA, 90048, USA.
⁶ School of Medicine, University of California San Diego, La Jolla, CA, 92093, USA.
⁷ Department of Ophthalmology, University of California San Diego, La Jolla, CA, 92093, USA. Electronic address: kangzhang@gmail.com.
⁸ Department of NanoEngineering, University of California San Diego, La Jolla, CA, 92093, USA; Materials Science and Engineering Program, University of California San Diego, La Jolla, CA, 92093, USA; Department of Bioengineering, University of California San Diego, La Jolla, CA, 92093, USA. Electronic address: chen168@eng.ucsd.edu.

Abstract

Ocular surface diseases including conjunctival disorders are multifactorial progressive conditions that can severely affect vision and quality of life. In recent years, stem cell therapies based on conjunctival stem cells (CjSCs) have become a potential solution for treating ocular surface diseases. However, neither an efficient culture of CjSCs nor the development of a minimally invasive ocular surface CjSC transplantation therapy has been reported. Here, we developed a robust in vitro expansion method for primary rabbit-derived CjSCs and applied digital light processing (DLP)-based bioprinting to produce CjSC-loaded hydrogel micro-constructs for injectable delivery. Expansion medium containing small molecule cocktail generated fast dividing and highly homogenous CjSCs for more than 10 passages in feeder-free culture. Bioprinted hydrogel micro-constructs with tunable mechanical properties enabled the 3D culture of CjSCs while supporting viability, stem cell phenotype, and differentiation potency into conjunctival goblet cells. These hydrogel micro-constructs were well-suited for scalable dynamic suspension culture of CjSCs and were successfully delivered to the bulbar conjunctival epithelium via minimally invasive subconjunctival injection. This work integrates novel cell culture strategies with bioprinting to develop a clinically relevant injectable-delivery approach for CjSCs towards the stem cell therapies for the treatment of ocular surface diseases.

Keywords: 3D culture; Bioprinting; Conjunctival stem cell; Injectable delivery; Ocular surface regeneration; Stem cell therapy.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Bioprinting*
Cell Differentiation
Hydrogels
Printing, Three-Dimensional
Quality of Life
Rabbits
Stem Cells
Tissue Engineering

Substances

Hydrogels

Abstract

Publication types

MeSH terms

Substances

Grants and funding