Dewi, Sianty, Tjahjono, Yudy, Novita, Bernadette Dian, Wijaya, Hendy, Putra, Brilliant Dwi, Widodo, Teguh, Hendriati, Lucia, Jong, FX. Himawan Haryanto, Malonda, Franklin V. and Kuncorojakti, Suryo (2025) Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum. Universa Medicina, 44 (1). pp. 65-72. ISSN P-ISSN : 19073062, E-ISSN : 24072230
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Abstract
BACKGROUND Mesenchymal stem cells (MSCs) are valued in regenerative medicine for their multipotency, proliferative capacity, and immunomodulatory properties. Wharton’s jelly-derived MSCs (WJ-MSCs) from the umbilical cord offer a non-invasive, promising source for clinical applications, because easy isolation, lack of ethical concerns, and the presence of both embryonic and adult stem cells have made them a valuable source for use in therapeutic applications and regenerative medicine. This study aimed to optimize WJ-MSC isolation and characterization methods. METHODS Human umbilical cords from three healthy donors were collected post-cesarean under strict inclusion criteria. WJ-MSCs were isolated using the explant culture method, with cells adhering to T75 flasks pre-coated with 2% gelatin. Cultures were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% freshly prepared Human AB serum and monitored for 21 days. Flow cytometry (BD FACSAria) was performed at passages 1 and 5 to assess MSC markers CD105, CD73, CD90, and CD44, alongside the exclusion marker CD45. RESULTS WJ-MSCs exhibited fibroblast-like morphology by passage 1 and showed robust proliferation. Flow cytometry revealed high CD44 expression (~60%) at passage 1, while CD105, CD73, and CD90 became prominent by passage 5. CD45 remained low, suggesting minimal hematopoietic contamination. CONCLUSIONS This study confirms the feasibility of isolating and expanding WJ-MSCs using DMEM with 10% human AB serum. While consistent cell growth was achieved, the 21-day culture period may require optimization for scalability, including serum concentration, substrate coatings, and oxygen levels. CPJ-MSCs may be preferable for applications demanding rapid expansion and early marker expression.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Wharton’s jelly, mesenchymal stem cells, isolation, flow cytometry, regenerative medicine |
| Subjects: | Medicine |
| Divisions: | Journal Publication |
| Depositing User: | F.X. Hadi |
| Date Deposited: | 13 Aug 2025 04:14 |
| Last Modified: | 13 Aug 2025 04:14 |
| URI: | https://repositori.ukwms.ac.id/id/eprint/44377 |
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