TY - JOUR
T1 - Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix
AU - Ro, Hyunuk
AU - Park, Jungha
AU - Yang, Kisuk
AU - Kim, Jiyong
AU - Yim, Hyun Gu
AU - Jung, Giyoung
AU - Lee, Hyukjin
AU - Cho, Seung Woo
AU - Hwang, Nathaniel S.
N1 - Funding Information:
This research was supported by the Basic Science Research Program (grant no. 0458–20120013) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning (MSIP). This study was also partially supported by a grant (2009–0083522) from the Translational Research Center for Protein Function Control (TRCP) funded by the Ministry of Science, ICT, and Future Planning (MSIP), Republic of Korea.
Publisher Copyright:
© 2015, Springer-Verlag Berlin Heidelberg.
PY - 2015/10/22
Y1 - 2015/10/22
N2 - Transient cartilage and a mineralizing microenvironment play pivotal roles in mesenchymal cell ossification during bone formation. In order to recreate these microenvironmental cues, C3H10T1/2 murine mesenchymal stem cells (MSCs) were exposed to chondrocyte-conditioned medium (CM) and seeded onto three-dimensional mineralized scaffolds for bone regeneration. Expansion of C3H10T1/2 cells with CM resulted in enhanced expression levels of chondrogenic markers such as aggrecan, type II collagen, type X collagen, and Sox9, rather than of osteogenic genes. Interestingly, CM expansion led to reduced expression levels of osteogenic genes such as alkaline phosphatase (ALP), type I collagen, osteocalcin, and Runx2. However, CM-expanded C3H10T1/2 cells showed enhanced osteogenic differentiation as indicated by increased ALP and Alizarin Red S staining upon osteogenic factor exposure. In vivo, CM-expanded C3H10T1/2 mesenchymal cells were seeded onto mineralized scaffolds (fabricated with polydopamine and coated with simulated body fluids) and implanted into critical-sized calvarial-defect mouse models. After 8 weeks of implantation, mouse skulls were collected, and bone tissue regeneration was evaluated by micro-computed tumography and Masson’s trichrome staining. In accordance with the in vitro analysis, CM-expanded C3H10T1/2 cells gave enhanced bone mineral deposition. Thus, chondrocyte-conditioned factors and a mineralized microenvironment stimulate the bone formation of MSCs.
AB - Transient cartilage and a mineralizing microenvironment play pivotal roles in mesenchymal cell ossification during bone formation. In order to recreate these microenvironmental cues, C3H10T1/2 murine mesenchymal stem cells (MSCs) were exposed to chondrocyte-conditioned medium (CM) and seeded onto three-dimensional mineralized scaffolds for bone regeneration. Expansion of C3H10T1/2 cells with CM resulted in enhanced expression levels of chondrogenic markers such as aggrecan, type II collagen, type X collagen, and Sox9, rather than of osteogenic genes. Interestingly, CM expansion led to reduced expression levels of osteogenic genes such as alkaline phosphatase (ALP), type I collagen, osteocalcin, and Runx2. However, CM-expanded C3H10T1/2 cells showed enhanced osteogenic differentiation as indicated by increased ALP and Alizarin Red S staining upon osteogenic factor exposure. In vivo, CM-expanded C3H10T1/2 mesenchymal cells were seeded onto mineralized scaffolds (fabricated with polydopamine and coated with simulated body fluids) and implanted into critical-sized calvarial-defect mouse models. After 8 weeks of implantation, mouse skulls were collected, and bone tissue regeneration was evaluated by micro-computed tumography and Masson’s trichrome staining. In accordance with the in vitro analysis, CM-expanded C3H10T1/2 cells gave enhanced bone mineral deposition. Thus, chondrocyte-conditioned factors and a mineralized microenvironment stimulate the bone formation of MSCs.
KW - Conditioned medium
KW - In vivo implantation
KW - Mineralized scaffolds
KW - Mouse (BALB/c)
KW - Osteogenic differentiation
KW - Simulated body fluids
KW - Stem cells
KW - Tissue engineering
UR - http://www.scopus.com/inward/record.url?scp=84941877298&partnerID=8YFLogxK
U2 - 10.1007/s00441-015-2195-7
DO - 10.1007/s00441-015-2195-7
M3 - Article
C2 - 25956591
AN - SCOPUS:84941877298
VL - 362
SP - 115
EP - 126
JO - Cell and Tissue Research
JF - Cell and Tissue Research
SN - 0302-766X
IS - 1
ER -