The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis
Developing fibrochondrogenic serum-free media is important for regenerating diseased and injured fibrocartilage but no defined protocols exist. Towards this goal, we characterized the effect of four candidate fibrochondrogenic serum-free media containing transforming growth factor beta-3 (TGF-β3), i...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Bioengineering |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2306-5354/12/5/546 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849711249874485248 |
|---|---|
| author | Ka Yu Carissa Kwan Ke Li Yu Yang Wang Wai Yi Tse Chung Yan Tong Xu Zhang Dan Michelle Wang Dai Fei Elmer Ker |
| author_facet | Ka Yu Carissa Kwan Ke Li Yu Yang Wang Wai Yi Tse Chung Yan Tong Xu Zhang Dan Michelle Wang Dai Fei Elmer Ker |
| author_sort | Ka Yu Carissa Kwan |
| collection | DOAJ |
| description | Developing fibrochondrogenic serum-free media is important for regenerating diseased and injured fibrocartilage but no defined protocols exist. Towards this goal, we characterized the effect of four candidate fibrochondrogenic serum-free media containing transforming growth factor beta-3 (TGF-β3), insulin-like growth factor-1 (IGF-1), and fibroblast growth factor-2 (FGF-2) with high/low glucose and with/without dexamethasone on human mesenchymal stem cells (hMSCs) via proliferation and differentiation assays. In Ki67 proliferation assays, serum-free media containing low glucose and dexamethasone exhibited the highest growth. In gene expression assays, serum-free media containing low glucose and commercially available chondrogenic media (COM) induced high fibrochondrogenic transcription factor expression (scleraxis/<i>SCX</i> and SRY-Box Transcription Factor 9/<i>SOX9</i>) and extracellular matrix (ECM) protein levels (aggrecan/<i>ACAN,</i> collagen type I/<i>COL1A1</i>, and collagen type II/<i>COL2A1</i>), respectively. In immunofluorescence staining, serum-free media containing high glucose and COM induced high fibrochondrogenic transcription factor (SCX and SOX9) and ECM protein (COL1A1, COL2A1, and collagen type X/COL10A1) levels, respectively. In cytochemical staining, COM and serum-free media containing dexamethasone showed a high collagen content whereas serum-free media containing high glucose and dexamethasone exhibited high glycosaminoglycan (GAG) levels. Altogether, defined serum-free media containing high glucose exhibited the highest fibrochondrogenic potential. In summary, this work studied conditions conducive for fibrochondrogenesis, which may be further optimized for potential applications in fibrocartilage tissue engineering. |
| format | Article |
| id | doaj-art-45b3f7e2ad9943c0a1cf862c5eeab37a |
| institution | DOAJ |
| issn | 2306-5354 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj-art-45b3f7e2ad9943c0a1cf862c5eeab37a2025-08-20T03:14:40ZengMDPI AGBioengineering2306-53542025-05-0112554610.3390/bioengineering12050546The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell FibrochondrogenesisKa Yu Carissa Kwan0Ke Li1Yu Yang Wang2Wai Yi Tse3Chung Yan Tong4Xu Zhang5Dan Michelle Wang6Dai Fei Elmer Ker7School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaSchool of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, ChinaDepartment of Biomedical Engineering, Faculty of Engineering, The Hong Kong Polytechnic University, Hung Hom, Hong Kong SAR, ChinaDeveloping fibrochondrogenic serum-free media is important for regenerating diseased and injured fibrocartilage but no defined protocols exist. Towards this goal, we characterized the effect of four candidate fibrochondrogenic serum-free media containing transforming growth factor beta-3 (TGF-β3), insulin-like growth factor-1 (IGF-1), and fibroblast growth factor-2 (FGF-2) with high/low glucose and with/without dexamethasone on human mesenchymal stem cells (hMSCs) via proliferation and differentiation assays. In Ki67 proliferation assays, serum-free media containing low glucose and dexamethasone exhibited the highest growth. In gene expression assays, serum-free media containing low glucose and commercially available chondrogenic media (COM) induced high fibrochondrogenic transcription factor expression (scleraxis/<i>SCX</i> and SRY-Box Transcription Factor 9/<i>SOX9</i>) and extracellular matrix (ECM) protein levels (aggrecan/<i>ACAN,</i> collagen type I/<i>COL1A1</i>, and collagen type II/<i>COL2A1</i>), respectively. In immunofluorescence staining, serum-free media containing high glucose and COM induced high fibrochondrogenic transcription factor (SCX and SOX9) and ECM protein (COL1A1, COL2A1, and collagen type X/COL10A1) levels, respectively. In cytochemical staining, COM and serum-free media containing dexamethasone showed a high collagen content whereas serum-free media containing high glucose and dexamethasone exhibited high glycosaminoglycan (GAG) levels. Altogether, defined serum-free media containing high glucose exhibited the highest fibrochondrogenic potential. In summary, this work studied conditions conducive for fibrochondrogenesis, which may be further optimized for potential applications in fibrocartilage tissue engineering.https://www.mdpi.com/2306-5354/12/5/546fibrocartilage tissue engineeringhuman mesenchymal stem cellsserum-free mediumgrowth factorsglucosedexamethasone |
| spellingShingle | Ka Yu Carissa Kwan Ke Li Yu Yang Wang Wai Yi Tse Chung Yan Tong Xu Zhang Dan Michelle Wang Dai Fei Elmer Ker The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis Bioengineering fibrocartilage tissue engineering human mesenchymal stem cells serum-free medium growth factors glucose dexamethasone |
| title | The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis |
| title_full | The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis |
| title_fullStr | The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis |
| title_full_unstemmed | The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis |
| title_short | The Characterization of Serum-Free Media on Human Mesenchymal Stem Cell Fibrochondrogenesis |
| title_sort | characterization of serum free media on human mesenchymal stem cell fibrochondrogenesis |
| topic | fibrocartilage tissue engineering human mesenchymal stem cells serum-free medium growth factors glucose dexamethasone |
| url | https://www.mdpi.com/2306-5354/12/5/546 |
| work_keys_str_mv | AT kayucarissakwan thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT keli thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT yuyangwang thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT waiyitse thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT chungyantong thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT xuzhang thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT danmichellewang thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT daifeielmerker thecharacterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT kayucarissakwan characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT keli characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT yuyangwang characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT waiyitse characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT chungyantong characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT xuzhang characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT danmichellewang characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis AT daifeielmerker characterizationofserumfreemediaonhumanmesenchymalstemcellfibrochondrogenesis |