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Fibroblast growth factor-2 maintains the differentiation potential of nucleus pulposus cells in vitro: implications for cell-based transplantation therapy.
Spine (Phila Pa 1976). 2007 Mar 01; 32(5):495-502.S

Abstract

STUDY DESIGN

To investigate effects of FGF-2 on nucleus pulposus cell growth and differentiation.

OBJECTIVES

To elucidate the phenotypic changes that occur during expansion of nucleus pulposus cells in monolayer culture, and to investigate the effects of fibroblast growth factor (FGF)-2 on cell growth and differentiation.

SUMMARY OF BACKGROUND DATA

Nucleus pulposus cells would have a limited application for autologous cell transplantation if phenotypic dedifferentiation takes place during culture expansion. FGF-2 has been shown to retain the differentiation potential of monolayer expanded chondrocytic cells. However, its effect on nucleus pulposus cells is not known.

METHODS

Bovine nucleus pulposus cells were serially passaged in the presence or absence of FGF-2 (1 and 10 ng/mL). After passage numbers 1 and 7, cells were immobilized in alginate beads and treated with transforming growth factor (TGF)-beta1 for 1 week to assess their differentiation.

RESULTS

During culture expansion in monolayer, nucleus pulposus cells maintained the expression of aggrecan messenger ribonucleic acid (mRNA). However, mRNA levels of collagen type I, collagen type II, Sox-9, and versican decreased with increasing passage number for both control (untreated) cells and FGF-2 treated cells. When grown in alginate with TFG-beta1, passage 7 cells that received FGF-2 during culture expansion restored the mRNA expression of type II collagen, Sox-9, COMP, chondroadherin, and fibromodulin. Moreover, FGF-2 treatment resulted in increased sulfated proteoglycan synthesis and lower aggrecan turnover compared to untreated controls under identical culture conditions. FGF-2 treated cells continued to express HIF-1alpha protein till passage 7, while MMP-2 expression was evident in cells treated with TGF-beta1. In addition, cells pretreated with FGF-2 showed higher induction of phospho ERK1/2 after treatment with TGF-beta1. Also, FGF-2 maintained smad 2/smad 3 mediated signaling in cells after TGF-beta treatment. FGF-2 action resulted in reduced actin stress fiber formation and migratory cell morphology, with no effect on cell proliferation.

CONCLUSIONS

The presence of FGF-2 during culture expansion of nucleus pulposus cells in monolayer can sustain a differentiated cell phenotype by maintaining responsiveness to TGF-beta1. Our results suggest that FGF-2 should be tested for its ability to maintain the reactivity of the nucleus pulposus cells to other morphogenic factors that may be used for cell-based transplantation therapy.

Authors+Show Affiliations

Department of Orthopaedic Surgery and Graduate Program in Tissue Engineering and Regenerative Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

17334282

Citation

Tsai, Tsung-Ting, et al. "Fibroblast Growth Factor-2 Maintains the Differentiation Potential of Nucleus Pulposus Cells in Vitro: Implications for Cell-based Transplantation Therapy." Spine, vol. 32, no. 5, 2007, pp. 495-502.
Tsai TT, Guttapalli A, Oguz E, et al. Fibroblast growth factor-2 maintains the differentiation potential of nucleus pulposus cells in vitro: implications for cell-based transplantation therapy. Spine (Phila Pa 1976). 2007;32(5):495-502.
Tsai, T. T., Guttapalli, A., Oguz, E., Chen, L. H., Vaccaro, A. R., Albert, T. J., Shapiro, I. M., & Risbud, M. V. (2007). Fibroblast growth factor-2 maintains the differentiation potential of nucleus pulposus cells in vitro: implications for cell-based transplantation therapy. Spine, 32(5), 495-502.
Tsai TT, et al. Fibroblast Growth Factor-2 Maintains the Differentiation Potential of Nucleus Pulposus Cells in Vitro: Implications for Cell-based Transplantation Therapy. Spine (Phila Pa 1976). 2007 Mar 1;32(5):495-502. PubMed PMID: 17334282.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Fibroblast growth factor-2 maintains the differentiation potential of nucleus pulposus cells in vitro: implications for cell-based transplantation therapy. AU - Tsai,Tsung-Ting, AU - Guttapalli,Asha, AU - Oguz,Erbil, AU - Chen,Lih-Huei, AU - Vaccaro,Alexander R, AU - Albert,Todd J, AU - Shapiro,Irving M, AU - Risbud,Makarand V, PY - 2007/3/6/pubmed PY - 2007/3/24/medline PY - 2007/3/6/entrez SP - 495 EP - 502 JF - Spine JO - Spine (Phila Pa 1976) VL - 32 IS - 5 N2 - STUDY DESIGN: To investigate effects of FGF-2 on nucleus pulposus cell growth and differentiation. OBJECTIVES: To elucidate the phenotypic changes that occur during expansion of nucleus pulposus cells in monolayer culture, and to investigate the effects of fibroblast growth factor (FGF)-2 on cell growth and differentiation. SUMMARY OF BACKGROUND DATA: Nucleus pulposus cells would have a limited application for autologous cell transplantation if phenotypic dedifferentiation takes place during culture expansion. FGF-2 has been shown to retain the differentiation potential of monolayer expanded chondrocytic cells. However, its effect on nucleus pulposus cells is not known. METHODS: Bovine nucleus pulposus cells were serially passaged in the presence or absence of FGF-2 (1 and 10 ng/mL). After passage numbers 1 and 7, cells were immobilized in alginate beads and treated with transforming growth factor (TGF)-beta1 for 1 week to assess their differentiation. RESULTS: During culture expansion in monolayer, nucleus pulposus cells maintained the expression of aggrecan messenger ribonucleic acid (mRNA). However, mRNA levels of collagen type I, collagen type II, Sox-9, and versican decreased with increasing passage number for both control (untreated) cells and FGF-2 treated cells. When grown in alginate with TFG-beta1, passage 7 cells that received FGF-2 during culture expansion restored the mRNA expression of type II collagen, Sox-9, COMP, chondroadherin, and fibromodulin. Moreover, FGF-2 treatment resulted in increased sulfated proteoglycan synthesis and lower aggrecan turnover compared to untreated controls under identical culture conditions. FGF-2 treated cells continued to express HIF-1alpha protein till passage 7, while MMP-2 expression was evident in cells treated with TGF-beta1. In addition, cells pretreated with FGF-2 showed higher induction of phospho ERK1/2 after treatment with TGF-beta1. Also, FGF-2 maintained smad 2/smad 3 mediated signaling in cells after TGF-beta treatment. FGF-2 action resulted in reduced actin stress fiber formation and migratory cell morphology, with no effect on cell proliferation. CONCLUSIONS: The presence of FGF-2 during culture expansion of nucleus pulposus cells in monolayer can sustain a differentiated cell phenotype by maintaining responsiveness to TGF-beta1. Our results suggest that FGF-2 should be tested for its ability to maintain the reactivity of the nucleus pulposus cells to other morphogenic factors that may be used for cell-based transplantation therapy. SN - 1528-1159 UR - https://www.unboundmedicine.com/medline/citation/17334282/Fibroblast_growth_factor_2_maintains_the_differentiation_potential_of_nucleus_pulposus_cells_in_vitro:_implications_for_cell_based_transplantation_therapy_ L2 - https://doi.org/10.1097/01.brs.0000257341.88880.f1 DB - PRIME DP - Unbound Medicine ER -