J Vet Sci.  2015 Sep;16(3):381-384. 10.4142/jvs.2015.16.3.381.

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

• Souza TF
• Sakamoto S
• Ferreira G
• Gameiro R
• Marinho M
• de Andrade AL
• Cardoso TC

• KMID: 2344312
• DOI: http://doi.org/10.4142/jvs.2015.16.3.381

Abstract

Canine mesenchymal cells (MSCs) derived from Wharton's jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.

Keyword

bone; mesenchymal stem cells; platelet-rich plasma; umbilical cord

MeSH Terms

Animals
Bone Demineralization Technique/veterinary
Bone Matrix/*metabolism
Cell Differentiation
Cells, Cultured
Coculture Techniques/veterinary
Dogs
Mesenchymal Stromal Cells/*metabolism
*Osteogenesis
Platelet-Rich Plasma/*metabolism
Umbilical Cord/metabolism

Figure

• 

  Fig. 1 (A) Fibroblast-like morphology of canine MSCs isolated Wharton's jelly at the 10th passage. Micrographs were taken under phase contrast microscopy. 40× magnification. MSCs phenotyping by flow cytometer analysis: (B) CD34-/CD44+, (C) CD45-/CD271+, (D) CD45-/CD105+. Data from surface markers were obtained by histograms generated using an Attune acoustic focusing cytometer, and auto-fluorescence was excluded as a global compensation tool (cutoff > 104). Y-axis corresponds to the number of positive labeled cells detected by BL1A filter (488 nm).

• 

  Fig. 2 (A) Alizarin Red staining showing extracellular matrix deposition and mineralization. 40× magnification. Alkaline phosphatase levels (405 nm; Y-axis) from control (MSCs)β, MSCs + PRP, MSCs + DBM, and MSCs + PRP + DBM groups at 0, 7, 14 and 21 days after culture (*p < 0.05). (B) Osteopontin, osteocalcin and Runx2 transcription in all studied groups at 0, 7, 14 and 21 days after culture.

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