In vitro culture of osteoblasts without the use of animal derived material
-
1
Cardiff University, Department of Child Health, School of Medicine, United Kingdom
-
2
Cardiff University, Division of Pathophysiology and Repair, School of Biosciences, United Kingdom
-
3
Cardiff University, Tissue Engineering and Reparative Dentistry, School of Dentistry, United Kingdom
It is important to advance and implement the 3Rs i.e. replacing, reducing and refining the use of animals in research and testing. Furthermore, musculoskeletal cell or tissue replacement therapies require the use of chemically defined media, which do not contain animal derived materials. We investigated the use of a range of non-FCS containing media on cell number (3 days; MTS assay) in human osteoblast lines (MG63, SaOS2). We have compared the current gold standard of DMEM containing 10% FCS with a range of test media: 1) DMEM with 10% human serum (Lonza), 2) DMEM with 10% human serum AB (Lonza) 3) DMEM with K/O serum replacement (10%, 15%; Invitrogen), and 4) TheraPEAK (defined medium; Lonza). We also investigated the effects of these media on multipotent, marrow-derived, human mesenchymal stem cell (MSC; Promocell) numbers and differentiation (3 days; ALP activity). Cells were either i) set up in control media and test media added the next day, or ii) set up in the test media.{BR}With MG63 cells, human serum and K/O decreased (50-60%; p<0.001), whereas TheraPEAK increased numbers (~130%; p<0.001). With the more differentiated SaOS2, however, human serum increased (120%; p<0.01) or had no effect, depending on the set up methodology used, K/O (15%) decreased (~35%; p<0.01), whereas TheraPEAK increased numbers by 200 to 300% (p<0.001). With undifferentiated MSCs, results were similar to those with MG63 cells, although reductions with human serum and K/O were ~25% and ~75% respectively (p<0.001). Human serum and TheraPEAK stimulated (~250%; p<0.001) differentiation (ALP activity) in MSCs. Human bone chips set up in different media resulted in the establishment of osteoblast cultures in all media, although FCS containing cultures exhibited most proliferation, whereas osteoblasts in human serum containing medium were the most differentiated. Very few cells were observed with TheraPEAK.{BR}The work shows that cells at different stages of the osteoblast differentiation pathway respond differently to non-FCS containing media, and that it is possible to establish primary, human osteoblast cultures without using FCS. The results will guide our planned studies to design successful osteoblast and osteocyte culture methods using media which do not contain animal derived materials.
Keywords:
Bones,
Bone Research
Conference:
2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society, Cambridge, United Kingdom, 27 Jun - 29 Jun, 2011.
Presentation Type:
Poster
Topic:
Abstracts
Citation:
Elford
C,
Mason
D,
Ralphs
J,
Gregory
J,
Sloan
A and
Evans
B
(2011). In vitro culture of osteoblasts without the use of animal derived material.
Front. Endocrinol.
Conference Abstract:
2011 joint meeting of the Bone Research Society & the British Orthopaedic Research Society.
doi: 10.3389/conf.fendo.2011.02.00017
Copyright:
The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers.
They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.
The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated.
Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed.
For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions.
Received:
30 Sep 2011;
Published Online:
30 Sep 2011.
*
Correspondence:
Dr. BAJ Evans, Cardiff University, Department of Child Health, School of Medicine, United Kingdom, evansba@cardiff.ac.uk