|ITEM METADATA RECORD
|Title: ||Pre-culture under serum free conditions enhances the osteochondrogenic response of human periosteum derived cells to BMP-2|
|Authors: ||Bolander, Johanna ×|
Luyten, Frank #
|Issue Date: ||17-Sep-2014 |
|Host Document: ||Pre-culture under serum free conditions enhances the osteochondrogenic response of human periosteum derived cells to BMP-2|
|Conference: ||International BMP Confernece edition:10 location:Berlin date:16-20th September 2014|
|Abstract: ||Combining osteoinductive biomolecules, osteochondroprogenitor cells and osteoconductive carriers into an engineered construct, may be a therapeutic strategy to successfully heal large critical bone defects. Bone morphogenetic protein-2 (BMP-2) is a potent osteoinductive soluble molecule based on its in vitro and in vivo biological activity. With respect to osteochondroprogenitor cells, human periosteum-derived cells (hPDCs) are promising progenitor cells in view of their direct contribution to postnatal fracture healing. Standard in vitro expansion of hPDCs involves media supplemented with 10% fetal bovine serum (FBS). However, the use of FBS equates to xenogeneic antigens and zoonosis contaminations, thus brings a potential impediment for future clinical translation. Therefore, the aim of this study was to investigate the effect of pre-culture followed by BMP-2 stimulation of hPDCs under serum free conditions in vitro and in vivo.
To this purpose, hPDC proliferation was monitored during a 21-day culture period in media containing different serum levels. A serum free chemically defined medium (CDM) was found to effectively maintain hPDC viability without inducing significant proliferation. Furthermore, an enhanced osteochondrogenic differentiation profile was seen of hPDCs in response to BMP-2 supplemented CDM evidenced by a 2.5-, 6-, 5- and 4-fold increased expression of Sox9, Aggrecan, Runx2 and Osterix. Culture media containing 10 % FBS was used as standard control. Therefore, CDM was further selected to pre-culture hPDCs for 6 days followed by 6 days of stimulation with BMP-2 supplemented CDM. Mesenchymal stem cell phenotype, proliferation, and osteochondrogenic differentiation were analysed. Pre-culture in CDM also resulted in a more than 2-fold increase in expression of the BMP receptors ALK1, ALK2, ALK3, ALK6, BMPR2, AcvR2a and AcvR2b. Furthermore, a 1.5-fold increase in ALP-activity was detected in hPDCs after pre-culture in CDM followed by BMP-2 stimulation. On gene expression level, enhanced osteochondrogenic differentiation was displayed by an increase of 2-, 1.5-, 2.5- and 3-fold expression of Sox9, Aggrecan, Runx2 and Osteocalcin, respectively. This could be correlated to activated BMP-signalling, verified by an increased expression of ID1. Surprisingly, immunohistochemistry for Sox9 and Osterix indicated that a fraction of the cells expressed both markers.
Together, these data display that an enhanced effect of BMP-2 on osteochondrogenic cell proliferation, differentiation and downstream BMP signalling can be achieved when serum containing medium is replaced by a serum free chemically defined medium. These findings provide an opportunity for the preparation of clinically relevant cell based constructs for in vivo bone tissue regeneration.
|Publication status: ||published|
|KU Leuven publication type: ||IMa|
|Appears in Collections:||Skeletal Biology and Engineering Research Center (+)|
Materials Technology TC, Campus Group T Leuven
Department of Materials Engineering - miscellaneous
× corresponding author|
# (joint) last author|
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