Collagen-Derived Dipeptide Pro-Hyp Enhanced ATDC5 Chondrocyte Differentiation under Hypoxic Conditions
Abstract
:1. Introduction
2. Results and Discussion
2.1. Pro-Hyp Regulates Chondrogenic ATDC5 Cells Differentiation in Hypoxic Condition
2.2. Pro-Hyp Further Upregulates Chondrogenesis-Specific Genes and ECM Remodeling Regulators under Hypoxic Conditions Compared to Normoxic Conditions
2.3. Pro-Hyp Further Increases Sox9 Protein Level without Affecting Runx2 Protein Level under Hypoxic Conditions Compared to Normoxic Conditions
3. Materials and Methods
3.1. Chemicals
3.2. Cell Culture and Treatment
3.3. Cell Proliferation
3.4. Alcian Blue Staining
3.5. RNA Preparation and Quantitative RT-PCR (qPCR)
3.6. Western Blot Assay
3.7. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Forward Sequence (5′-3′) | Reverse Sequence (5′-3′) |
---|---|---|
Sox9 | CCAGCAAGAACAAGCCACAC | GCTCAGTTCACCGATGTCCA |
Col2α1 | AGGTGCTCAAGGTTCTCGTG | GCTCCAGGAAGACCAGGTTC |
Aggrecan | CCAAACCAGCCTGACAACTT | TCTAGCATGCTCCACCACTG |
Runx2 | TAAGAAGAGCCAGGAGGTGC | AGGTACGTGTGGTAGTGAGTG |
MMP9 | TGAATCAGCTGGCTTTTGTG | GTGGATAGCTCGGTGGTGTT |
MMP13 | AGGCCTTCAGAAAAGCCTTC | TCCTTGGAGTGATCCAGACC |
β-actin | AAGGCCAACCGTGAAAAGAT | GTGGTACGACCAGAGGCATAC |
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Kimira, Y.; Sato, T.; Sakamoto, M.; Osawa, Y.; Mano, H. Collagen-Derived Dipeptide Pro-Hyp Enhanced ATDC5 Chondrocyte Differentiation under Hypoxic Conditions. Molecules 2023, 28, 4664. https://doi.org/10.3390/molecules28124664
Kimira Y, Sato T, Sakamoto M, Osawa Y, Mano H. Collagen-Derived Dipeptide Pro-Hyp Enhanced ATDC5 Chondrocyte Differentiation under Hypoxic Conditions. Molecules. 2023; 28(12):4664. https://doi.org/10.3390/molecules28124664
Chicago/Turabian StyleKimira, Yoshifumi, Takahiro Sato, Mayu Sakamoto, Yoshihiro Osawa, and Hiroshi Mano. 2023. "Collagen-Derived Dipeptide Pro-Hyp Enhanced ATDC5 Chondrocyte Differentiation under Hypoxic Conditions" Molecules 28, no. 12: 4664. https://doi.org/10.3390/molecules28124664