gms | German Medical Science

130. Kongress der Deutschen Gesellschaft für Chirurgie

Deutsche Gesellschaft für Chirurgie

30.04. - 03.05.2013, München

The effect of different basal cell culture media during mesenchymal stem cell induction on their osteogenic differentiation potential after a DMEM high glucose based growth factor enhanced expansion in a standard osteogenic monolayer cell culture assayed with 99m-Tc-MDP Labeling

Meeting Abstract

Suche in Medline nach

  • Tobias Großner - Universitätsklinikum Heidelberg, Dept. Orthopädie und Unfallchirurgie, Heidelberg
  • Tobias Gotterbarm - Universitätsklinikum Heidelberg, Dept. Orthopädie und Unfallchirurgie, Heidelberg

Deutsche Gesellschaft für Chirurgie. 130. Kongress der Deutschen Gesellschaft für Chirurgie. München, 30.04.-03.05.2013. Düsseldorf: German Medical Science GMS Publishing House; 2013. Doc13dgch168

doi: 10.3205/13dgch168, urn:nbn:de:0183-13dgch1687

Veröffentlicht: 26. April 2013

© 2013 Großner et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielfältigt, verbreitet und öffentlich zugänglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Introduction: Maximizing the osteogenic potential of mesenchymal stem cells in vitro is of major interest in tissue engineering in order to maximize the bone formation. It is unclear if the use of different basal media during MSC induction significantly affects the amount of osteogenic differentiation and extracellular hydroxyapatite deposition.

Material and methods: Human MSCs (n=6) were thawed and 250.000 cells were expanded for 10 days using DMEM high glucose+MCDB201+2%FKS+ISTSupp.+1%P/S+0,02µMDexamethason+0,1µMAscorbicacid-2-phosphat+EGF 10 ng/ml + PDGF 10ng/ml. Then, cells were transferred for induction into 3,5cm diameter petri dishes (30.000 cells/cm2).

One dish from each donor was filled with one of five different basal cell culture media:

1) DMEM low glucose+10%FCS+1%P/S; 2) DMEM high glucose+10%FCS+1%P/S; 3) Alpha-MEM+10%FCS+1%P/S; 4) DMEM-F12+10%FCS+1%P/S+TGFß-1+FGF; 5) DMEM high glucose+MCDB201+2%FKS+ISTSupp.+1%P/S+0,02µMDexamethason+0,1µMAscorbicacid-2-phosphat+EGF 10 ng/ml+PDGF 10ng/ml (“Verfaillie”-Media). As osteogenic supplements were added: 10mM ß-glycerolphosphat+50µM Ascorbicacid2-phosphat+100nM Dexamethason.

Osteogenic induction was performed for 21 days and then terminated. 5,5 MBq 99mTc-MDP in 0,9%NaCl was added to each dish. After incubation dishes were placed under a gamma camera nd the gammacounts for each dish were acquired for 180 seconds.

SPSS 20 based ANOVA-Analysis with Post-Hoc-Test/Bonferroni-correction was performed to determine significant different amounts of osteogenic differentiation. P values of not more than 0.05 were considered as significant.

Results: We could show, that when DMEM-HG-based“Verfaillie”-Media is used for cell expansion followed by osteogenic induction, only media 1,3 and 4 show a significant osteogenic differentiation detected by radiated gamma counts. The highest uptake (=osteogenic differentiation) occurs when Alpha-MEM is used for induction (mean 43.531counts) compared to DMEM low glucose (mean 23.300counts) followed by 4/DMEM-F12 (mean 1604counts). We could determine a significant difference for Alpha-MEM compared to DMEM-F-12/+TGFß/FGF p=0,08.

No other significance were show which might be caused by a related in-group variability regarding the mineralization process.

Figure 1 [Fig. 1].

Conclusion: If using DMEM-HG-based“Verfaillie”-Media for cell-expansion media 1, 3 & 4 are suitable for osteogenic differentiation. We would favor Alpha-MEM for osteogenic induction.