Influence of perfusion and cyclic compression on proliferation and differentiation of bone marrow stromal cells in 3-dimensional culture

Jagodzinski, M.; Breitbart, A.; Wehmeier, M.; Hesse, E.; Haasper, C.; Krettek, C.; Zeichen, J.; Hankemeier, S.

doi:10.1016/j.jbiomech.2008.04.001

« Previous Next »Journal of Biomechanics
Volume 41, Issue 9 , Pages 1885-1891, 2008

Influence of perfusion and cyclic compression on proliferation and differentiation of bone marrow stromal cells in 3-dimensional culture

M. Jagodzinski
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
- Corresponding author. Tel.: +495115322278; fax: +495115325877.
A. Breitbart
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
M. Wehmeier
- Department of Clinical Chemistry, Hannover Medical School (MHH), Hanover, Germany
E. Hesse
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
- Current address: Department of Medicine, Harvard Medical School and Department of Oral Medicine, Infection and Immunity, Harvard School of Dental Medicine, Boston, MA, USA.
C. Haasper
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
C. Krettek
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
J. Zeichen
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany
S. Hankemeier
- Department of Trauma, Hannover Medical School (MHH), Carl-Neuberg-Street 1, D-30625 Hanover, Germany

Accepted 1 April 2008. published online 21 May 2008.

Abstract

Until now, there has been no in vitro model that duplicates the environment of bone marrow. The purpose of this study was to analyze proliferation and differentiation of human bone marrow stromal cells (hBMSC) under the influence of continuous perfusion and cyclic mechanical loading.

hBMSC of seven individuals were harvested, grown in vitro, and combined. 10⁶ hBMSC were seeded on a bovine spongiosa disc and incubated in a bioreactor system. Cell culture was continued using three different conditions: Continuous perfusion (group A), 10% cyclic compression at 0.5Hz (group B) and static controls (group C). After 24h, 1, 2, and 3 weeks, we determined cell proliferation (MTS-assay) and osteogenic differentiation (osteocalcin ELISA, Runx2 mRNA). Tenascin-C mRNA was quantified to exclude fibroblastic differentiation.

In groups A and B, proliferation was enhanced after 2 weeks (48.6±19.6×10³ (A) and 44.6±14.3×10³ cells (B)) and after 3 weeks (46.6±15.1×10³ (A) and 44.8±10.2×10³ cells (B)) compared with controls (26.3±10.8×10³ (2 weeks) and 17.1±6.5×10³ cells (3 weeks), p<0.03). Runx2 mRNA was upregulated in both stimulated groups after 1, 2, and 3 weeks compared to control (group A, 1 week: 5.2±0.7-fold; p<0.01, 2 weeks: 4.4±1.9-fold; p<0.01, 3 weeks: 3.8±1.7-fold; p=0.013; group B, 1 week: 3.6±1.1-fold, p<0.01, 2 weeks: 4.2±2.2-fold, p<0.01; 3 weeks: 5.3±2.7-fold, p<0.01). hBMSC stimulated by cyclic compression expressed the highest amount of osteocalcin at all time points (1 week: 294.5±88.4mg/g protein, 2 weeks: 294.4±73.3mg/g protein, 3 weeks: 293.1±83.6mg/g protein, p⩽0.03).

The main stimulus for cell proliferation in a 3-dimensional culture of hBMSC is continuous perfusion whereas mechanical stimulation fosters osteogenic commitment of hBMSC. This study thereby contributes to the understanding of physical stimuli that influence hBMSC in a 3-dimensional cell culture system.

Keywords: hBMSC, Bioreactor, Perfusion, Mechanical stimulation, Osteoblast differentiation