Gene expression profiles of early chondrogenic markers in dedifferentiated fat cells stimulated by bone morphogenetic protein 4 under monolayer and spheroid culture conditions in vitro. Issue 4 (1st December 2016)

• Record Type: Journal Article
• Authors: Azumi, Eiko; Honda, Yoshitomo; Kishimoto, Naotaka; Hashimoto, Yoshiya; Matsumoto, Naoyuki
• Is Part Of: Orthodontic waves. Volume 75:Issue 4(2016:Dec.)
• Page Start: 97
• Page End: 104
• Publication Date: 2016-12-01
• Subjects: Dedifferentiated fat cell -- Adipocyte -- Chondrocyte -- BMP-4 -- Differentiation; Orthodontics -- Periodicals; Orthodontics -- Periodicals; Orthodontie -- Périodiques; Orthodontie; Périodique électronique (Descripteur de forme); Ressource Internet (Descripteur de forme); 617.643
• DOI: 10.1016/j.odw.2016.10.006
• Languages: English
• ISSNs: 1344-0241

Abstract

Abstract: Purpose: Human dedifferentiated fat (hDFAT) cells are thought to be a promising cell source for cartilage regeneration therapy. Nevertheless, the responses of hDFAT cells to bone morphogenetic proteins (BMPs) are still unclear. Here, we elucidated the effects of BMP-4 on the mRNA expression of early chondrogenic markers in hDFAT cells under monolayer or pellet cell culture conditions. Materials and methods: Monolayer and pellet cell cultures of hDFAT cells were grown with control medium or chondrogenic medium (CM) with or without BMP-2, BMP-4, or BMP-7. Real-time polymerase chain reaction was used to analyze the mRNA expression levels of nine genes: chondrogenic markers, i.e., SOX9, SOX5, SOX6, aggrecan, type 2 collagen, type 10 collagen, and matrix metalloproteinase (MMP) 13; type 1 collagen; and MMP3 . The BMP signaling inhibitor dorsomorphin was used to verify the mechanisms of BMP-4-induced chondrogenesis. Results: Recombinant BMP-4 (100 ng/mL) increased the expression of SOX9, SOX6, and aggrecan mRNAs in monolayer cells compared with that in cells treated with BMP-2 or BMP-7 on day 3. Chondrogenically differentiated hDFAT cells induced by CM containing BMP-4 showed higher expression of eight genes (excluding SOX5 ) in monolayer cultures and nine genes (including SOX5 ) in pellet cultures compared with those in control medium on day 14. Dorsomorphin attenuated the effects of BMP-4. Conclusion: These results showed that BMP-4 had the potential to modulate the early chondrogenesis of hDFAT cells under both monolayer and pellet cell culture conditions.