Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice

Hiroyuki Nakahara, Haruo Misawa, Takahiro Hayashi, Eisaku Kondo, Takeshi Yuasa, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Wael A R A Hassan, Reham A R A Hassan, Shahid M. Javed, Masato Tanaka, Hirosuke Endo, Hirofumi Noguchi, Shinichi Matsumoto, Katsuyoshi Takata, Yuichi Tashiro, Shuhei Nakaji, Toshihumi Ozaki, Naoya Kobayashi

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

BACKGROUND. Human mesenchymal stem cells (hMSCs) are multipotent stem cells found in the adult bone marrow that have the capacity to differentiate into various mesenchymal cell types. The hMSCs may provide a potential therapy to restore damaged tissues or organs of mesenchymal origin; however, a drawback is their limited life span in vitro. METHODS. We immortalized normal hMSCs with retrovirally transmitted human telomerase reverse transcriptase cDNA. One of the immortalized clones (YKNK-12) was established, and the biological characteristics were investigated in vitro and in vivo. RESULTS. YKNK-12 cells were capable of differentiating adipocytes, osetoblasts, and chondrocytes. Osteogenically differentiated YKNK-12 cells produced significant levels of growth factors BMP4, BMP6, FGF6, FGF7, transforming growth factor-β1, and transforming growth factor-β3.. Microcomputer tomography T and soft X-ray assays showed an excellent calvarial bone healing in mice after transplantation of osteogenically differentiated YKNK-12 cells. These cells expressed human-specific osteocalcin and increased the gene expression of runt-related transcription factor 2, alkaline phosphatase, osteocalcin, and osterix in the bone regenerating area. YKNK-12 cell transplant corrected the bone defect without inducing any adverse effects. CONCLUSIONS. We conclude that hMSCs immortalized by transduction with human telomerase reverse transcriptase may provide an unlimited source of cells for therapeutic use in bone regeneration.

Original languageEnglish
Pages (from-to)346-353
Number of pages8
JournalTransplantation
Volume88
Issue number3
DOIs
Publication statusPublished - Aug 15 2009

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Bone Transplantation
Mesenchymal Stromal Cells
Osteocalcin
Transforming Growth Factors
Bone and Bones
Multipotent Stem Cells
Bone Regeneration
Microcomputers
Therapeutic Uses
Chondrocytes
Adipocytes
Alkaline Phosphatase
Intercellular Signaling Peptides and Proteins
Transcription Factors
Complementary DNA
Clone Cells
Transplantation
Bone Marrow
Tomography
X-Rays

Keywords

  • Bone marrow
  • hTERT
  • Mesenchymal stem cells

ASJC Scopus subject areas

  • Transplantation

Cite this

Nakahara, H., Misawa, H., Hayashi, T., Kondo, E., Yuasa, T., Kubota, Y., ... Kobayashi, N. (2009). Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice. Transplantation, 88(3), 346-353. https://doi.org/10.1097/TP.0b013e3181ae5ba2

Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice. / Nakahara, Hiroyuki; Misawa, Haruo; Hayashi, Takahiro; Kondo, Eisaku; Yuasa, Takeshi; Kubota, Yasuhiro; Seita, Masayuki; Kawamoto, Hironobu; Hassan, Wael A R A; Hassan, Reham A R A; Javed, Shahid M.; Tanaka, Masato; Endo, Hirosuke; Noguchi, Hirofumi; Matsumoto, Shinichi; Takata, Katsuyoshi; Tashiro, Yuichi; Nakaji, Shuhei; Ozaki, Toshihumi; Kobayashi, Naoya.

In: Transplantation, Vol. 88, No. 3, 15.08.2009, p. 346-353.

Research output: Contribution to journalArticle

Nakahara, H, Misawa, H, Hayashi, T, Kondo, E, Yuasa, T, Kubota, Y, Seita, M, Kawamoto, H, Hassan, WARA, Hassan, RARA, Javed, SM, Tanaka, M, Endo, H, Noguchi, H, Matsumoto, S, Takata, K, Tashiro, Y, Nakaji, S, Ozaki, T & Kobayashi, N 2009, 'Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice', Transplantation, vol. 88, no. 3, pp. 346-353. https://doi.org/10.1097/TP.0b013e3181ae5ba2
Nakahara, Hiroyuki ; Misawa, Haruo ; Hayashi, Takahiro ; Kondo, Eisaku ; Yuasa, Takeshi ; Kubota, Yasuhiro ; Seita, Masayuki ; Kawamoto, Hironobu ; Hassan, Wael A R A ; Hassan, Reham A R A ; Javed, Shahid M. ; Tanaka, Masato ; Endo, Hirosuke ; Noguchi, Hirofumi ; Matsumoto, Shinichi ; Takata, Katsuyoshi ; Tashiro, Yuichi ; Nakaji, Shuhei ; Ozaki, Toshihumi ; Kobayashi, Naoya. / Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice. In: Transplantation. 2009 ; Vol. 88, No. 3. pp. 346-353.
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AU - Nakahara, Hiroyuki

AU - Misawa, Haruo

AU - Hayashi, Takahiro

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AU - Yuasa, Takeshi

AU - Kubota, Yasuhiro

AU - Seita, Masayuki

AU - Kawamoto, Hironobu

AU - Hassan, Wael A R A

AU - Hassan, Reham A R A

AU - Javed, Shahid M.

AU - Tanaka, Masato

AU - Endo, Hirosuke

AU - Noguchi, Hirofumi

AU - Matsumoto, Shinichi

AU - Takata, Katsuyoshi

AU - Tashiro, Yuichi

AU - Nakaji, Shuhei

AU - Ozaki, Toshihumi

AU - Kobayashi, Naoya

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N2 - BACKGROUND. Human mesenchymal stem cells (hMSCs) are multipotent stem cells found in the adult bone marrow that have the capacity to differentiate into various mesenchymal cell types. The hMSCs may provide a potential therapy to restore damaged tissues or organs of mesenchymal origin; however, a drawback is their limited life span in vitro. METHODS. We immortalized normal hMSCs with retrovirally transmitted human telomerase reverse transcriptase cDNA. One of the immortalized clones (YKNK-12) was established, and the biological characteristics were investigated in vitro and in vivo. RESULTS. YKNK-12 cells were capable of differentiating adipocytes, osetoblasts, and chondrocytes. Osteogenically differentiated YKNK-12 cells produced significant levels of growth factors BMP4, BMP6, FGF6, FGF7, transforming growth factor-β1, and transforming growth factor-β3.. Microcomputer tomography T and soft X-ray assays showed an excellent calvarial bone healing in mice after transplantation of osteogenically differentiated YKNK-12 cells. These cells expressed human-specific osteocalcin and increased the gene expression of runt-related transcription factor 2, alkaline phosphatase, osteocalcin, and osterix in the bone regenerating area. YKNK-12 cell transplant corrected the bone defect without inducing any adverse effects. CONCLUSIONS. We conclude that hMSCs immortalized by transduction with human telomerase reverse transcriptase may provide an unlimited source of cells for therapeutic use in bone regeneration.

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KW - Bone marrow

KW - hTERT

KW - Mesenchymal stem cells

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