Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells
Abstract Background Lubricin, a proteoglycan encoded by the PRG4 gene, is synthesised by superficial zone (SFZ) chondrocytes and synovial cells. It reduces friction between joints and allows smooth sliding of tendons. Although lubricin has been shown to be effective against osteoarthritis and synovi...
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| Format: | Article |
| Language: | English |
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BMC
2022-07-01
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| Series: | BMC Molecular and Cell Biology |
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| Online Access: | https://doi.org/10.1186/s12860-022-00431-8 |
| _version_ | 1817968759314317312 |
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| author | Takashi Satake Shingo Komura Hitomi Aoki Akihiro Hirakawa Yuuki Imai Haruhiko Akiyama |
| author_facet | Takashi Satake Shingo Komura Hitomi Aoki Akihiro Hirakawa Yuuki Imai Haruhiko Akiyama |
| author_sort | Takashi Satake |
| collection | DOAJ |
| description | Abstract Background Lubricin, a proteoglycan encoded by the PRG4 gene, is synthesised by superficial zone (SFZ) chondrocytes and synovial cells. It reduces friction between joints and allows smooth sliding of tendons. Although lubricin has been shown to be effective against osteoarthritis and synovitis in animals, its clinical application remains untested. In this study, we aimed to induce lubricin-expressing cells from pluripotent stem cells (iPSCs) and applied them locally via cell transplantation. Methods To generate iPSCs, OCT3/4, SOX2, KLF4, and L-MYC were transduced into fibroblasts derived from Prg4-mRFP1 transgenic mice. We established a protocol for the differentiation of iPSC-derived Prg4-mRFP1-positive cells and characterised their mRNA expression profile. Finally, we injected Prg4-mRFP1-positive cells into the paratenon, surrounding the Achilles tendons and knee joints of severe combined immunodeficient mice and assessed lubricin expression. Result Wnt3a, activin A, TGF-β1, and bFGF were applied to induce the differentiation of iPSC-derived Prg4-mRFP1-positive cells. Markers related to SFZ chondrocytes and fibroblast-like synovial cells (FLSs) were expressed during differentiation. RNA-sequencing indicated that iPSC-derived Prg4-mRFP1-positive cells manifested expression profiles typical of SFZ chondrocytes and FLSs. Transplanted iPSC-derived Prg4-mRFP1-positive cells survived around the Achilles tendons and in knee joints. Conclusions The present study describes a protocol for the differentiation of iPSC-derived Prg4-positive cells with characteristics of SFZ chondrocytes and FLSs. Transplantation of lubricin-expressing cells offers promise as a therapy against arthritis and synovitis. |
| first_indexed | 2024-04-13T20:12:05Z |
| format | Article |
| id | doaj.art-ac8de62031834471a9ccb077a3e3de4c |
| institution | Directory Open Access Journal |
| issn | 2661-8850 |
| language | English |
| last_indexed | 2024-04-13T20:12:05Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj.art-ac8de62031834471a9ccb077a3e3de4c2022-12-22T02:31:48ZengBMCBMC Molecular and Cell Biology2661-88502022-07-0123111310.1186/s12860-022-00431-8Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cellsTakashi Satake0Shingo Komura1Hitomi Aoki2Akihiro Hirakawa3Yuuki Imai4Haruhiko Akiyama5Department of Orthopaedic Surgery, Gifu University Graduate School of MedicineDepartment of Orthopaedic Surgery, Gifu University Graduate School of MedicineDepartment of Tissue and Organ Development, Advanced Medical Science, Gifu University Graduate School of MedicineDepartment of Orthopaedic Surgery, Gifu University Graduate School of MedicineDivision of Integrative Pathophysiology, Proteo-Science Center, Graduate School of Medicine, Ehime UniversityDepartment of Orthopaedic Surgery, Gifu University Graduate School of MedicineAbstract Background Lubricin, a proteoglycan encoded by the PRG4 gene, is synthesised by superficial zone (SFZ) chondrocytes and synovial cells. It reduces friction between joints and allows smooth sliding of tendons. Although lubricin has been shown to be effective against osteoarthritis and synovitis in animals, its clinical application remains untested. In this study, we aimed to induce lubricin-expressing cells from pluripotent stem cells (iPSCs) and applied them locally via cell transplantation. Methods To generate iPSCs, OCT3/4, SOX2, KLF4, and L-MYC were transduced into fibroblasts derived from Prg4-mRFP1 transgenic mice. We established a protocol for the differentiation of iPSC-derived Prg4-mRFP1-positive cells and characterised their mRNA expression profile. Finally, we injected Prg4-mRFP1-positive cells into the paratenon, surrounding the Achilles tendons and knee joints of severe combined immunodeficient mice and assessed lubricin expression. Result Wnt3a, activin A, TGF-β1, and bFGF were applied to induce the differentiation of iPSC-derived Prg4-mRFP1-positive cells. Markers related to SFZ chondrocytes and fibroblast-like synovial cells (FLSs) were expressed during differentiation. RNA-sequencing indicated that iPSC-derived Prg4-mRFP1-positive cells manifested expression profiles typical of SFZ chondrocytes and FLSs. Transplanted iPSC-derived Prg4-mRFP1-positive cells survived around the Achilles tendons and in knee joints. Conclusions The present study describes a protocol for the differentiation of iPSC-derived Prg4-positive cells with characteristics of SFZ chondrocytes and FLSs. Transplantation of lubricin-expressing cells offers promise as a therapy against arthritis and synovitis.https://doi.org/10.1186/s12860-022-00431-8iPSCsPrg4Superficial zone chondrocytesFibroblast-like synovial cells |
| spellingShingle | Takashi Satake Shingo Komura Hitomi Aoki Akihiro Hirakawa Yuuki Imai Haruhiko Akiyama Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells BMC Molecular and Cell Biology iPSCs Prg4 Superficial zone chondrocytes Fibroblast-like synovial cells |
| title | Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells |
| title_full | Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells |
| title_fullStr | Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells |
| title_full_unstemmed | Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells |
| title_short | Induction of iPSC-derived Prg4-positive cells with characteristics of superficial zone chondrocytes and fibroblast-like synovial cells |
| title_sort | induction of ipsc derived prg4 positive cells with characteristics of superficial zone chondrocytes and fibroblast like synovial cells |
| topic | iPSCs Prg4 Superficial zone chondrocytes Fibroblast-like synovial cells |
| url | https://doi.org/10.1186/s12860-022-00431-8 |
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