gms | German Medical Science

Deutscher Kongress für Orthopädie und Unfallchirurgie
74. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie
96. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie
51. Tagung des Berufsverbandes der Fachärzte für Orthopädie und Unfallchirurgie

26. - 29.10.2010, Berlin

Adipose-derived Mesenchymal Stem Cells spontaneously differentiate into Osteoblast-like cells

Meeting Abstract

  • I. Kerler - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany
  • S. Ehnert - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany
  • A. Schmitt - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany
  • M. Lucke - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany
  • U. Stöckle - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany
  • A. K. Nussler - Klinikum Rechts der Isar der TU München, Unfallchirurgie, München, Germany

Deutscher Kongress für Orthopädie und Unfallchirurgie. 74. Jahrestagung der Deutschen Gesellschaft für Unfallchirurgie, 96. Tagung der Deutschen Gesellschaft für Orthopädie und Orthopädische Chirurgie, 51. Tagung des Berufsverbandes der Fachärzte für Orthopädie. Berlin, 26.-29.10.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. DocIN23-1326

DOI: 10.3205/10dkou145, URN: urn:nbn:de:0183-10dkou1455

Published: October 21, 2010

© 2010 Kerler et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.


Outline

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Objective: The use of Mesenchymal Stem Cells (MSCs) for the tissue engineering of vascularized bone is widely spread because of their ability to differentiate towards both required cell types - bone and epithelial cells. Bone Marrow was always thought to be one of the most promising sources of MSCs. However the isolation of Bone Marrow bares some risks for the patient because of the invasive extraction method and the unpredictable effects on the patients haematopoetic balance. Whereas Adipose derived MSCs (Ad-MSCs) represent a more easily available and less dangerous source of such adult stem cells. Aim of this project was to improve the osteogenic differentiation of Ad-MSCs.

Methods: Osteoblasts and B-MSCs were isolated from femur heads of patients undergoing prosthetic replacement and Ad-MSCs were isolated from wasted hip or abdominal fat tissue of patients, in accordance to the ethical vote of the TU Munich. We checked presence of surface receptors and osteogenic markers by RT-PCR. BMP signaling was checked by adenoviral reporter assays. Formation of mineralized matrix was confirmed by von Kossa and alizarin red staining and AP activity was measured photometrically.

Results and conclusions: The proliferation rate of Ad-MSCs was comparable to B-MSCs. Ad-MSCs express receptors for BMPs (Alk1, -2, -3, -5, -6, TGF-βRII & BMP-RII), vitamin D, insulin (IGF-R I & II) and Retinol (receptor β &γ). Only very high concentrations of BMP2 and/or BMP7 induced Smad1/5/8 signaling but not Smad2/3 signaling in MSCs. Based on this characterization we prepared and optimized our osteogenic differentiation medium. Supplementation of medium with β-glycerol-phosphate, L-ascorbic-acid phosphate, CaCl2 and vitamin D results in an increase of AP activity comparable to primary human osteoblasts (20–160 μmol/min) already after 8 days. Using the same medium, between 12 to 16 days the matrix mineralization started. At the same time expression of osteoblastic markers such as BMP2, osteopontin and osteocalcin began. Further addition of high conc. of BMP2 and/or BMP7 did not significantly improve the osteogenic differentiation.

Our results show, that Ad-MSCs have great potential for osteogenic differentiation, even without addition of cytokines, and thus represent an excellent alternative for B-MSCs in tissue engineering.