Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation

Sun-Ae Oh, Hye-Young Lee, Jae Ho Lee, Tae-Hyun Kim, Jun-Hyeog Jang, Hae-Won Kim, Ivan Wall

Research output: Contribution to journalArticle

Abstract

Three-dimensional (3D) collagen hydrogels have been extensively used for cell culture experiments and are more closely representative of in vivo conditions than monolayer (2D) culture. Here we cultured rat bone marrow-derived mesenchymal stem cells (MSCs) in collagen hydrogels containing varying concentrations of basic fibroblast growth factor (bFGF) to examine the effect of bFGF on MSC proliferation and osteogenic differentiation in 3D culture. The optimal bFGF concentration that promoted the greatest degree of cell proliferation and expression of the early osteogenic induction marker alkaline phosphatase was also determined. Subsequent quantitative real-time polymerase chain reaction analysis of gene expression demonstrated that bFGF promoted significant upregulation of the bone-related genes: collagen type I, osteopontin (OPN), bone sialoprotein (BSP), and osteocalcin (OCN) for periods of up to 21 days. Immunofluorescence staining and fluorescence-activated cell sorting analysis further supported the enhanced osteogenic differentiation of cells as a greater proportion of cells were found to express OPN. Matrix mineralization within the collagen hydrogels was enhanced in the presence of bFGF, as assessed by calcium detection using von Kossa staining. These results clearly demonstrate a positive effect of bFGF on proliferation and osteogenic induction of MSCs in 3D collagen hydrogels when applied at the appropriate concentration. Moreover, collagen hydrogel constructs containing MSCs and appropriate growth factor stimulus might be a potentially useful biological tool for 3D bone tissue engineering.

Original languageEnglish
Pages (from-to)1087-100
Number of pages14
JournalTissue Engineering: Parts A, B and C
Volume18
Issue number9-10
DOIs
Publication statusPublished - May 2012

Fingerprint

Hydrogel
Fibroblast Growth Factor 2
Mesenchymal Stromal Cells
Cell Differentiation
Hydrogels
Collagen
Osteopontin
Cell Proliferation
Integrin-Binding Sialoprotein
Staining and Labeling
Bone and Bones
Osteocalcin
Tissue Engineering
Collagen Type I
Fluorescent Antibody Technique
Alkaline Phosphatase
Real-Time Polymerase Chain Reaction
Intercellular Signaling Peptides and Proteins
Flow Cytometry
Up-Regulation

Keywords

  • Animals
  • Blotting, Western
  • Cell Differentiation/drug effects
  • Cell Proliferation/drug effects
  • Cells, Cultured
  • Collagen/chemistry
  • Fibroblast Growth Factor 2/chemistry
  • Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry
  • Male
  • Mesenchymal Stem Cells
  • Osteogenesis/drug effects
  • Polymerase Chain Reaction
  • Rats
  • Rats, Sprague-Dawley

Cite this

Oh, Sun-Ae ; Lee, Hye-Young ; Lee, Jae Ho ; Kim, Tae-Hyun ; Jang, Jun-Hyeog ; Kim, Hae-Won ; Wall, Ivan. / Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation. In: Tissue Engineering: Parts A, B and C. 2012 ; Vol. 18, No. 9-10. pp. 1087-100.
@article{691c94a70a3544c0961251d3a8d2e5ab,
title = "Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation",
abstract = "Three-dimensional (3D) collagen hydrogels have been extensively used for cell culture experiments and are more closely representative of in vivo conditions than monolayer (2D) culture. Here we cultured rat bone marrow-derived mesenchymal stem cells (MSCs) in collagen hydrogels containing varying concentrations of basic fibroblast growth factor (bFGF) to examine the effect of bFGF on MSC proliferation and osteogenic differentiation in 3D culture. The optimal bFGF concentration that promoted the greatest degree of cell proliferation and expression of the early osteogenic induction marker alkaline phosphatase was also determined. Subsequent quantitative real-time polymerase chain reaction analysis of gene expression demonstrated that bFGF promoted significant upregulation of the bone-related genes: collagen type I, osteopontin (OPN), bone sialoprotein (BSP), and osteocalcin (OCN) for periods of up to 21 days. Immunofluorescence staining and fluorescence-activated cell sorting analysis further supported the enhanced osteogenic differentiation of cells as a greater proportion of cells were found to express OPN. Matrix mineralization within the collagen hydrogels was enhanced in the presence of bFGF, as assessed by calcium detection using von Kossa staining. These results clearly demonstrate a positive effect of bFGF on proliferation and osteogenic induction of MSCs in 3D collagen hydrogels when applied at the appropriate concentration. Moreover, collagen hydrogel constructs containing MSCs and appropriate growth factor stimulus might be a potentially useful biological tool for 3D bone tissue engineering.",
keywords = "Animals, Blotting, Western, Cell Differentiation/drug effects, Cell Proliferation/drug effects, Cells, Cultured, Collagen/chemistry, Fibroblast Growth Factor 2/chemistry, Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry, Male, Mesenchymal Stem Cells, Osteogenesis/drug effects, Polymerase Chain Reaction, Rats, Rats, Sprague-Dawley",
author = "Sun-Ae Oh and Hye-Young Lee and Lee, {Jae Ho} and Tae-Hyun Kim and Jun-Hyeog Jang and Hae-Won Kim and Ivan Wall",
year = "2012",
month = "5",
doi = "10.1089/ten.TEA.2011.0360",
language = "English",
volume = "18",
pages = "1087--100",
journal = "Tissue Engineering: Parts A, B and C",
issn = "2152-4947",
publisher = "Mary Ann Liebert Inc.",
number = "9-10",

}

Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation. / Oh, Sun-Ae; Lee, Hye-Young; Lee, Jae Ho; Kim, Tae-Hyun; Jang, Jun-Hyeog; Kim, Hae-Won; Wall, Ivan.

In: Tissue Engineering: Parts A, B and C, Vol. 18, No. 9-10, 05.2012, p. 1087-100.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Collagen three-dimensional hydrogel matrix carrying basic fibroblast growth factor for the cultivation of mesenchymal stem cells and osteogenic differentiation

AU - Oh, Sun-Ae

AU - Lee, Hye-Young

AU - Lee, Jae Ho

AU - Kim, Tae-Hyun

AU - Jang, Jun-Hyeog

AU - Kim, Hae-Won

AU - Wall, Ivan

PY - 2012/5

Y1 - 2012/5

N2 - Three-dimensional (3D) collagen hydrogels have been extensively used for cell culture experiments and are more closely representative of in vivo conditions than monolayer (2D) culture. Here we cultured rat bone marrow-derived mesenchymal stem cells (MSCs) in collagen hydrogels containing varying concentrations of basic fibroblast growth factor (bFGF) to examine the effect of bFGF on MSC proliferation and osteogenic differentiation in 3D culture. The optimal bFGF concentration that promoted the greatest degree of cell proliferation and expression of the early osteogenic induction marker alkaline phosphatase was also determined. Subsequent quantitative real-time polymerase chain reaction analysis of gene expression demonstrated that bFGF promoted significant upregulation of the bone-related genes: collagen type I, osteopontin (OPN), bone sialoprotein (BSP), and osteocalcin (OCN) for periods of up to 21 days. Immunofluorescence staining and fluorescence-activated cell sorting analysis further supported the enhanced osteogenic differentiation of cells as a greater proportion of cells were found to express OPN. Matrix mineralization within the collagen hydrogels was enhanced in the presence of bFGF, as assessed by calcium detection using von Kossa staining. These results clearly demonstrate a positive effect of bFGF on proliferation and osteogenic induction of MSCs in 3D collagen hydrogels when applied at the appropriate concentration. Moreover, collagen hydrogel constructs containing MSCs and appropriate growth factor stimulus might be a potentially useful biological tool for 3D bone tissue engineering.

AB - Three-dimensional (3D) collagen hydrogels have been extensively used for cell culture experiments and are more closely representative of in vivo conditions than monolayer (2D) culture. Here we cultured rat bone marrow-derived mesenchymal stem cells (MSCs) in collagen hydrogels containing varying concentrations of basic fibroblast growth factor (bFGF) to examine the effect of bFGF on MSC proliferation and osteogenic differentiation in 3D culture. The optimal bFGF concentration that promoted the greatest degree of cell proliferation and expression of the early osteogenic induction marker alkaline phosphatase was also determined. Subsequent quantitative real-time polymerase chain reaction analysis of gene expression demonstrated that bFGF promoted significant upregulation of the bone-related genes: collagen type I, osteopontin (OPN), bone sialoprotein (BSP), and osteocalcin (OCN) for periods of up to 21 days. Immunofluorescence staining and fluorescence-activated cell sorting analysis further supported the enhanced osteogenic differentiation of cells as a greater proportion of cells were found to express OPN. Matrix mineralization within the collagen hydrogels was enhanced in the presence of bFGF, as assessed by calcium detection using von Kossa staining. These results clearly demonstrate a positive effect of bFGF on proliferation and osteogenic induction of MSCs in 3D collagen hydrogels when applied at the appropriate concentration. Moreover, collagen hydrogel constructs containing MSCs and appropriate growth factor stimulus might be a potentially useful biological tool for 3D bone tissue engineering.

KW - Animals

KW - Blotting, Western

KW - Cell Differentiation/drug effects

KW - Cell Proliferation/drug effects

KW - Cells, Cultured

KW - Collagen/chemistry

KW - Fibroblast Growth Factor 2/chemistry

KW - Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry

KW - Male

KW - Mesenchymal Stem Cells

KW - Osteogenesis/drug effects

KW - Polymerase Chain Reaction

KW - Rats

KW - Rats, Sprague-Dawley

UR - https://www.liebertpub.com/doi/10.1089/ten.tea.2011.0360

U2 - 10.1089/ten.TEA.2011.0360

DO - 10.1089/ten.TEA.2011.0360

M3 - Article

C2 - 22145747

VL - 18

SP - 1087

EP - 1100

JO - Tissue Engineering: Parts A, B and C

JF - Tissue Engineering: Parts A, B and C

SN - 2152-4947

IS - 9-10

ER -