Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix

Hyunuk Ro, Jungha Park, Kisuk Yang, Jiyong Kim, Hyun Gu Yim, Giyoung Jung, Hyukjin Lee, Seung Woo Cho, Nathaniel S. Hwang

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


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.

Original languageEnglish
Pages (from-to)115-126
Number of pages12
JournalCell and Tissue Research
Issue number1
StatePublished - 22 Oct 2015


  • Conditioned medium
  • In vivo implantation
  • Mineralized scaffolds
  • Mouse (BALB/c)
  • Osteogenic differentiation
  • Simulated body fluids
  • Stem cells
  • Tissue engineering


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