Fabrication of cartilage tissue substitutes from cells with induced pluripotency

About authors

1 Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia

2 Lomonosov Moscow State University, Moscow, Russia

3 Koltzov Institute of Developmental Biology, Moscow, Russia

Correspondence should be addressed: Artem V. Eremeev
Malaya Pirogovskaya, 1a, 119435, Moscow, Russia; ur.xednay@veemere-tra

About paper

Funding: the study was supported with an allocation #22-15-00250 by the Russian Science Foundation.

Acknowledgments: the authors thank Corresponding Member of the Russian Academy of Sciences M.A. Lagarkova for providing research facilities for the study

Author contribution: Eremeev AV — design of the experiment, general guidance, article authoring; Pikina AS — literature review, collection of the material, participation in the experimental part of the work, analysis of the resulting data; Ruchko ES — participation in the experimental part of the work; Sidorov VS, Ragozin AO — provision of material for the experiment.

Compliance with ethical standards: the study was performed in conformity with the principles of the Declaration of Helsinki (2000) and its subsequent revisions.

Received: 2022-10-12 Accepted: 2022-10-28 Published online: 2022-11-23

One of the approaches to cartilage tissue restoration problem relies on cellular technologies that use iPSCs, induced pluripotency stem cells that are an unlimited source of cellular material for tissue engineering with significant differentiation potential. However, there are no standardized protocols for chondrogenic differentiation of iPSCs. This study aimed to make cartilage tissue samples using 3D spheroid cultures and following four chondrogenic differentiation protocols, then compare characteristics of the cartilage samples made under different protocols and isolate the most effective way of differentiation. The iPSCs were differentiated chondrogenically, the four protocols were "long", "short", "combined" and with conditioned medium from a primary culture of autologous chondrocytes; the combinations of TGFβ1, BMP2, Chir 99021, and PK factors varied. Microwell plates were used to make spheroids. Immunocytochemical staining, real-time PCR and histological staining enabled assessment of the synthesis and expression profiles. High rates of synthesis and expression of chondrogenic markers Sox9, aggrecan, type II collagen were observed in spheroids experimented with under the "long", "combined" protocols and the conditioned medium protocol. The "combined" differentiation protocol made chondrogenesis most effective, and conditioned medium was highly efficient in inducing and supporting chondrogenic differentiation.

Keywords: tissue engineering, articular cartilage, induced pluripotency stem cells (iPSCs), spheroids, chondrogenesis