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Generation of scaffoldless hyaline cartilaginous tissue from human iPSCs.

Yamashita A, Morioka M, Yahara Y, Okada M, Kobayashi T, Kuriyama S, Matsuda S, Tsumaki N - Stem Cell Reports (2015)

Bottom Line: Defects in articular cartilage ultimately result in loss of joint function.The immunodeficiency mice and rats suffered from neither tumors nor ectopic tissue formation.The hiPSC-derived cartilaginous particles constitute a viable cell source for regenerating cartilage defects.

View Article: PubMed Central - PubMed

Affiliation: Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.

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Orthotopic Transplantation of hiPSC-Derived Cells into SCID RatshiPSC-derived cartilaginous particles obtained on day 28 were transplanted into defects created in the articular cartilage of the distal femurs of SCID rats. The transplanted sites (A and B) and various organs (C) were collected.(A and B) Histological analysis of the transplanted sites at 1 and 4 weeks after transplantation. Semiserial sections were stained with H&E and toluidine blue and immunostained with anti-vimentin antibodies that recognize only human vimentin and anti-type II collagen antibodies. The blue color reflects DAPI. Magnified images of the boxed regions in (A) are shown in (B). Scale bars, 50 μm.(C) RNAs were extracted from various organs at 4 and 12 weeks after transplantation and subjected to real-time RT-PCR to amplify human and rat β-actin mRNAs. n = 3 rats. The error bars denote the means ± SD. Bone, bone of the femoral diaphysis; Surrounding fat, fat tissue surrounding the transplanted sites; Intraperitoneal, intraperitoneal tissue; Groin, groin lymph nodes; Axillary, axillary lymph nodes; Cervical, cervical lymph nodes; MEF, murine embryonic fibroblasts; HDF, human dermal fibroblasts.See also Figure S6 and Tables S1–S3.
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fig6: Orthotopic Transplantation of hiPSC-Derived Cells into SCID RatshiPSC-derived cartilaginous particles obtained on day 28 were transplanted into defects created in the articular cartilage of the distal femurs of SCID rats. The transplanted sites (A and B) and various organs (C) were collected.(A and B) Histological analysis of the transplanted sites at 1 and 4 weeks after transplantation. Semiserial sections were stained with H&E and toluidine blue and immunostained with anti-vimentin antibodies that recognize only human vimentin and anti-type II collagen antibodies. The blue color reflects DAPI. Magnified images of the boxed regions in (A) are shown in (B). Scale bars, 50 μm.(C) RNAs were extracted from various organs at 4 and 12 weeks after transplantation and subjected to real-time RT-PCR to amplify human and rat β-actin mRNAs. n = 3 rats. The error bars denote the means ± SD. Bone, bone of the femoral diaphysis; Surrounding fat, fat tissue surrounding the transplanted sites; Intraperitoneal, intraperitoneal tissue; Groin, groin lymph nodes; Axillary, axillary lymph nodes; Cervical, cervical lymph nodes; MEF, murine embryonic fibroblasts; HDF, human dermal fibroblasts.See also Figure S6 and Tables S1–S3.

Mentions: We transplanted hiPSC-derived cartilaginous particles into defects created in the articular cartilage of SCID rats. Due to the small size and limited depth of the rat cartilage, we were unable to fix mature particles that were lubricious in their defects. Therefore, we transplanted premature-hiPSC-derived cartilaginous particles obtained on day 28. The defects were filled with hiPSC-derived cells in three of four knees at 1 week and three of four knees at 4 weeks after transplantation, as indicated by the expression of human vimentin (Figure 6A). The day-28 particles produced tissue that exhibited metachromatic staining with toluidine blue and a strong expression of type II collagen in the articular cartilage defects (Figures 6A and 6B), which differs from the observation that day-28 particles fail to produce mature cartilage in subcutaneous spaces. We speculate that the orthotopic environment might stimulate the maturation of day-28 particles. Side-to-side integration between the tissues formed by the transplanted cells and the rat articular cartilage was strongly achieved (Figure 6B). There were no signs of teratomas or other tumors in any of the four transplanted sites.


Generation of scaffoldless hyaline cartilaginous tissue from human iPSCs.

Yamashita A, Morioka M, Yahara Y, Okada M, Kobayashi T, Kuriyama S, Matsuda S, Tsumaki N - Stem Cell Reports (2015)

Orthotopic Transplantation of hiPSC-Derived Cells into SCID RatshiPSC-derived cartilaginous particles obtained on day 28 were transplanted into defects created in the articular cartilage of the distal femurs of SCID rats. The transplanted sites (A and B) and various organs (C) were collected.(A and B) Histological analysis of the transplanted sites at 1 and 4 weeks after transplantation. Semiserial sections were stained with H&E and toluidine blue and immunostained with anti-vimentin antibodies that recognize only human vimentin and anti-type II collagen antibodies. The blue color reflects DAPI. Magnified images of the boxed regions in (A) are shown in (B). Scale bars, 50 μm.(C) RNAs were extracted from various organs at 4 and 12 weeks after transplantation and subjected to real-time RT-PCR to amplify human and rat β-actin mRNAs. n = 3 rats. The error bars denote the means ± SD. Bone, bone of the femoral diaphysis; Surrounding fat, fat tissue surrounding the transplanted sites; Intraperitoneal, intraperitoneal tissue; Groin, groin lymph nodes; Axillary, axillary lymph nodes; Cervical, cervical lymph nodes; MEF, murine embryonic fibroblasts; HDF, human dermal fibroblasts.See also Figure S6 and Tables S1–S3.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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fig6: Orthotopic Transplantation of hiPSC-Derived Cells into SCID RatshiPSC-derived cartilaginous particles obtained on day 28 were transplanted into defects created in the articular cartilage of the distal femurs of SCID rats. The transplanted sites (A and B) and various organs (C) were collected.(A and B) Histological analysis of the transplanted sites at 1 and 4 weeks after transplantation. Semiserial sections were stained with H&E and toluidine blue and immunostained with anti-vimentin antibodies that recognize only human vimentin and anti-type II collagen antibodies. The blue color reflects DAPI. Magnified images of the boxed regions in (A) are shown in (B). Scale bars, 50 μm.(C) RNAs were extracted from various organs at 4 and 12 weeks after transplantation and subjected to real-time RT-PCR to amplify human and rat β-actin mRNAs. n = 3 rats. The error bars denote the means ± SD. Bone, bone of the femoral diaphysis; Surrounding fat, fat tissue surrounding the transplanted sites; Intraperitoneal, intraperitoneal tissue; Groin, groin lymph nodes; Axillary, axillary lymph nodes; Cervical, cervical lymph nodes; MEF, murine embryonic fibroblasts; HDF, human dermal fibroblasts.See also Figure S6 and Tables S1–S3.
Mentions: We transplanted hiPSC-derived cartilaginous particles into defects created in the articular cartilage of SCID rats. Due to the small size and limited depth of the rat cartilage, we were unable to fix mature particles that were lubricious in their defects. Therefore, we transplanted premature-hiPSC-derived cartilaginous particles obtained on day 28. The defects were filled with hiPSC-derived cells in three of four knees at 1 week and three of four knees at 4 weeks after transplantation, as indicated by the expression of human vimentin (Figure 6A). The day-28 particles produced tissue that exhibited metachromatic staining with toluidine blue and a strong expression of type II collagen in the articular cartilage defects (Figures 6A and 6B), which differs from the observation that day-28 particles fail to produce mature cartilage in subcutaneous spaces. We speculate that the orthotopic environment might stimulate the maturation of day-28 particles. Side-to-side integration between the tissues formed by the transplanted cells and the rat articular cartilage was strongly achieved (Figure 6B). There were no signs of teratomas or other tumors in any of the four transplanted sites.

Bottom Line: Defects in articular cartilage ultimately result in loss of joint function.The immunodeficiency mice and rats suffered from neither tumors nor ectopic tissue formation.The hiPSC-derived cartilaginous particles constitute a viable cell source for regenerating cartilage defects.

View Article: PubMed Central - PubMed

Affiliation: Cell Induction and Regulation Field, Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.

Show MeSH
Related in: MedlinePlus