A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells

Sanne Karijn Both, Adrie J.C. van der Muijsenberg, Clemens van Blitterswijk, Jan de Boer, Joost Dick de Bruijn

Research output: Contribution to journalArticleAcademic

129 Citations (Scopus)

Abstract

During the past decade, there has been much interest in the use of human mesenchymal stem cells (hMSCs) in bone tissue engineering. HMSCs can be obtained relatively easily and expanded rapidly in culture, but for clinical purposes large numbers are often needed and the cost should be kept to a minimum. A rapid and efficient culturing protocol would therefore be beneficial. In this study, we examined the effect of different medium compositions on the expansion and osteogenic differentiation of bone marrow–derived hMSCs from 19 donors. We also investigated the effect of low seeding density and dexamethasone on both hMSCs expansion and their in vitro and in vivo osteogenic differentiation capacity. HMSCs seeded at a density of 100 cells/cm2 had a significantly higher growth rate than at 5000 cell/cm2, which was further improved by the addition of dexamethasone. Expanded hMSCs were characterized in vitro on the basis of positive staining for CD29, CD44, CD105, and CD166. The in vitro osteogenic potential of expanded hMSCs was assessed by flow cytometric staining for alkaline phosphatase. In vivo bone-forming potential of the hMSCs was assessed by seeding the cells in ceramic scaffolds, followed by subcutaneous implantation in nude mice and histopathologic assessment of de novo bone formation after 6-week implantation. Expanded hMSCs from all donors displayed similar osteogenic potential independent of the culture conditions. On the basis of these results we have developed an efficient method to culture hMSCs by seeding the cells at 100 cells/cm2 in an α-minimal essential medium–based medium containing dexamethasone.
Original languageUndefined
Pages (from-to)3-9
JournalTissue engineering
Volume13
Issue number1
DOIs
Publication statusPublished - 2007

Keywords

  • IR-67185

Cite this

Both, S. K., van der Muijsenberg, A. J. C., van Blitterswijk, C., de Boer, J., & de Bruijn, J. D. (2007). A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells. Tissue engineering, 13(1), 3-9. https://doi.org/10.1089/ten.2005.0513
Both, Sanne Karijn ; van der Muijsenberg, Adrie J.C. ; van Blitterswijk, Clemens ; de Boer, Jan ; de Bruijn, Joost Dick. / A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells. In: Tissue engineering. 2007 ; Vol. 13, No. 1. pp. 3-9.
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Both, SK, van der Muijsenberg, AJC, van Blitterswijk, C, de Boer, J & de Bruijn, JD 2007, 'A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells' Tissue engineering, vol. 13, no. 1, pp. 3-9. https://doi.org/10.1089/ten.2005.0513

A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells. / Both, Sanne Karijn; van der Muijsenberg, Adrie J.C.; van Blitterswijk, Clemens; de Boer, Jan; de Bruijn, Joost Dick.

In: Tissue engineering, Vol. 13, No. 1, 2007, p. 3-9.

Research output: Contribution to journalArticleAcademic

TY - JOUR

T1 - A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells

AU - Both, Sanne Karijn

AU - van der Muijsenberg, Adrie J.C.

AU - van Blitterswijk, Clemens

AU - de Boer, Jan

AU - de Bruijn, Joost Dick

PY - 2007

Y1 - 2007

N2 - During the past decade, there has been much interest in the use of human mesenchymal stem cells (hMSCs) in bone tissue engineering. HMSCs can be obtained relatively easily and expanded rapidly in culture, but for clinical purposes large numbers are often needed and the cost should be kept to a minimum. A rapid and efficient culturing protocol would therefore be beneficial. In this study, we examined the effect of different medium compositions on the expansion and osteogenic differentiation of bone marrow–derived hMSCs from 19 donors. We also investigated the effect of low seeding density and dexamethasone on both hMSCs expansion and their in vitro and in vivo osteogenic differentiation capacity. HMSCs seeded at a density of 100 cells/cm2 had a significantly higher growth rate than at 5000 cell/cm2, which was further improved by the addition of dexamethasone. Expanded hMSCs were characterized in vitro on the basis of positive staining for CD29, CD44, CD105, and CD166. The in vitro osteogenic potential of expanded hMSCs was assessed by flow cytometric staining for alkaline phosphatase. In vivo bone-forming potential of the hMSCs was assessed by seeding the cells in ceramic scaffolds, followed by subcutaneous implantation in nude mice and histopathologic assessment of de novo bone formation after 6-week implantation. Expanded hMSCs from all donors displayed similar osteogenic potential independent of the culture conditions. On the basis of these results we have developed an efficient method to culture hMSCs by seeding the cells at 100 cells/cm2 in an α-minimal essential medium–based medium containing dexamethasone.

AB - During the past decade, there has been much interest in the use of human mesenchymal stem cells (hMSCs) in bone tissue engineering. HMSCs can be obtained relatively easily and expanded rapidly in culture, but for clinical purposes large numbers are often needed and the cost should be kept to a minimum. A rapid and efficient culturing protocol would therefore be beneficial. In this study, we examined the effect of different medium compositions on the expansion and osteogenic differentiation of bone marrow–derived hMSCs from 19 donors. We also investigated the effect of low seeding density and dexamethasone on both hMSCs expansion and their in vitro and in vivo osteogenic differentiation capacity. HMSCs seeded at a density of 100 cells/cm2 had a significantly higher growth rate than at 5000 cell/cm2, which was further improved by the addition of dexamethasone. Expanded hMSCs were characterized in vitro on the basis of positive staining for CD29, CD44, CD105, and CD166. The in vitro osteogenic potential of expanded hMSCs was assessed by flow cytometric staining for alkaline phosphatase. In vivo bone-forming potential of the hMSCs was assessed by seeding the cells in ceramic scaffolds, followed by subcutaneous implantation in nude mice and histopathologic assessment of de novo bone formation after 6-week implantation. Expanded hMSCs from all donors displayed similar osteogenic potential independent of the culture conditions. On the basis of these results we have developed an efficient method to culture hMSCs by seeding the cells at 100 cells/cm2 in an α-minimal essential medium–based medium containing dexamethasone.

KW - IR-67185

U2 - 10.1089/ten.2005.0513

DO - 10.1089/ten.2005.0513

M3 - Article

VL - 13

SP - 3

EP - 9

JO - Tissue engineering

JF - Tissue engineering

SN - 1076-3279

IS - 1

ER -

Both SK, van der Muijsenberg AJC, van Blitterswijk C, de Boer J, de Bruijn JD. A Rapid and Efficient Method for Expansion of Human Mesenchymal Stem Cells. Tissue engineering. 2007;13(1):3-9. https://doi.org/10.1089/ten.2005.0513