Tissue engineering of the human corneal endothelium: From primary cell culture to transplantation of highly functional cell sheets

人角膜内皮的组织工程：从原代细胞培养到高功能细胞片的移植

• 批准号: 266879141
• 负责人: Professorin Dr. Katrin Engelmann
• 金额: --
• 依托单位: Leibniz-Institut für Polymerforschung Dresden (IPF)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/266879141?language=en
• 关键词: Tissue; engineering; human; corneal; endothelium

The proposed research project aims at in vitro cultivation and subsequent transfer of human corneal endothelial cell (HCEC) sheets for tissue engineering purposes. To allow a straightforward translation into a pre-clinical setting, primary, non-immortalized HCEC cultures are investigated. Isolation protocols and culture conditions are adapted to ensure an appropriate cell proliferation and maintenance. Highly functional and viable HCEC sheets are generated by methods which comply with essential requirements for later clinical application (e.g. animal compound free culture conditions). A tunable thermo-responsive cell culture carrier that was previously developed by the applicants and that allows to meet particular requirements of HCEC is employed. Upon stimulated detachment the cell sheets need to preserve intact intercellular contacts as well as extracellular matrix to stably survive transfer, storage and transplantation. Towards this goal a comprehensive range of characterization techniques for cell sheet morphology, viability, and physiological function is applied. Gentle manipulation techniques as well as dedicated transfer tools are developed using an in vitro transplantation model. More significant criteria for the transplant quality beyond the sole endothelial cell density are identified and evaluated.

拟议的研究项目旨在体外培养和随后转移用于组织工程目的的人角膜内皮细胞（HCEC）片。为了能够直接转化为临床前环境，对原代非永生化 HCEC 培养物进行了研究。调整分离方案和培养条件以确保适当的细胞增殖和维持。通过符合后期临床应用基本要求（例如无动物化合物培养条件）的方法生成高功能且可行的 HCEC 片。采用了申请人先前开发的并且允许满足HCEC的特定要求的可调热响应细胞培养载体。在受刺激的分离后，细胞片需要保留完整的细胞间接触以及细胞外基质，以在转移、储存和移植中稳定存活。为了实现这一目标，应用了细胞片形态、活力和生理功能的全面表征技术。使用体外移植模型开发了温和的操作技术以及专用的转移工具。确定并评估了除单一内皮细胞密度之外的更重要的移植质量标准。