Long-term repair of porcine articular cartilage using cryopreservable, clinically compatible human embryonic stem cell-derived chondrocytes
Osteoarthritis (OA) impacts hundreds of millions of people worldwide, with those affected incurring significant physical and financial burdens. Injuries such as focal defects to the articular surface are a major contributing risk factor for the development of OA. Current cartilage repair strategies are moderately effective at reducing pain but often replace damaged tissue with biomechanically inferior fibrocartilage. Here we describe the development, transcriptomic ontogenetic characterization and quality assessment at the single cell level, as well as the scaled manufacturing of an allogeneic human pluripotent stem cell-derived articular chondrocyte formulation that exhibits long-term functional repair of porcine articular cartilage. These results define a new potential clinical paradigm for articular cartilage repair and mitigation of the associated risk of OA.
Cell therapy has been used successfully in the clinic for more than 50 years in the form of hematopoietic stem cell transplantation1. This pioneering work illuminated the need for HLA-matched donors due to graft versus host disease (GVHD) encountered during allogenic transplants, in which donor lymphocytes reacted against host tissues. In the case of allogenic solid organ transplantation, immunosuppression of the host is often required for extended periods2. These aforementioned limitations in the availability and compatibility of donor tissue have prompted the search for other solutions which now potentially include human embryonic stem cell- (hESC) and induced pluripotent stem cell-derived (iPSC) cells and tissues3,4. The field of PSC-based regenerative medicine has advanced quickly as both iPSC- and ESC-derived cell therapies are in clinical trials5, with transplants into immunoprivileged sites such as the eye leading the way.
Scaled production and formulation optimization of hESC-derived chondrocytes
The main purpose of this study was to assess the long-term therapeutic potential of hESC-derived chondrocytes in a porcine model of focal articular cartilage injury. We have previously defined a protocol for the generation of articular cartilage-like chondrocytes from human PSCs17. Cells generated using this technique are immature based on their transcriptional signature and expression of immature chondrocyte markers but can mature upon implantation in vivo, evidencing appropriate expression of superficial zone markers and lack of hypertrophy18. For this study, we have adapted our previous protocols17,18 initially developed for H1 and H9 lines, to utilize the research grade hESC line ESI-01726. This specific line was selected because a cGMP version of this line is fully compliant with all current FDA regulations and can be advanced into human clinical trials without any regulatory restrictions.
In order to generate sufficient numbers of clinical grade hESC-derived chondrocytes for cartilage defect repair, hESCs were first expanded in hESC-qualified Matrigel and induced into mesodermal differentiation (d1–7) followed by chondrogenic differentiation (d7–11; Fig. 1a, Supplemental Materials).
Ref: https://www.nature.com/articles/s41536-021-00187-3
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