Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum
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...
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Faculty of Medicine Trisakti University
2025-04-01
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| Series: | Universa Medicina |
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| Online Access: | https://univmed.org/ejurnal/index.php/medicina/article/view/1678 |
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| author | Yudy Tjahjono Sianty Dewi Bernadette Dian Novita Hendy Wijaya Brilliant Dwi Putra Suryo Kuncorojakti Lucia Hendriati FX Himawan Haryanto Jong Teguh Widodo Franklin Vincentius Malonda |
| author_facet | Yudy Tjahjono Sianty Dewi Bernadette Dian Novita Hendy Wijaya Brilliant Dwi Putra Suryo Kuncorojakti Lucia Hendriati FX Himawan Haryanto Jong Teguh Widodo Franklin Vincentius Malonda |
| author_sort | Yudy Tjahjono |
| collection | DOAJ |
| description | 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.
Conclusion
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.
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| format | Article |
| id | doaj-art-a6b46bfece554335b2c4961fa578bc6e |
| institution | Kabale University |
| issn | 2407-2230 1907-3062 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Faculty of Medicine Trisakti University |
| record_format | Article |
| series | Universa Medicina |
| spelling | doaj-art-a6b46bfece554335b2c4961fa578bc6e2025-08-20T03:53:39ZengFaculty of Medicine Trisakti UniversityUniversa Medicina2407-22301907-30622025-04-0144110.18051/UnivMed.2025.v44.65-72Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serumYudy Tjahjono0https://orcid.org/0000-0003-0187-0270Sianty Dewi1https://orcid.org/0000-0003-1628-2615Bernadette Dian Novita2https://orcid.org/0000-0001-5624-7043Hendy Wijaya3https://orcid.org/0000-0003-2928-4791Brilliant Dwi Putra4Suryo Kuncorojakti5https://orcid.org/0000-0001-5170-9982Lucia Hendriati6https://orcid.org/0009-0000-0639-8759FX Himawan Haryanto Jong7https://orcid.org/0000-0002-3594-2327Teguh Widodo8https://orcid.org/0009-0005-4011-9043Franklin Vincentius Malonda9https://orcid.org/0000-0001-5170-9982Faculty of Medicine, Widya Mandala Catholic University Surabaya, East Java, Indonesia Faculty of Pharmacy, Widya Mandala Catholic University Surabaya, East Java, IndonesiaFaculty of Medicine, Widya Mandala Catholic University Surabaya, Surabaya, East Java, IndonesiaFaculty of Pharmacy, Widya Mandala Catholic University Surabaya, Surabaya, East Java, Indonesia Undergraduate Program, Faculty of Pharmacy, Widya Mandala Catholic University Surabaya, Surabaya, East Java, IndonesiaResearch Centre for Vaccine Technology and Development, Institute of Tropical Disease, Universitas Airlangga, Surabaya, IndonesiaFaculty of Pharmacy, Widya Mandala Catholic University Surabaya, Surabaya, East Java, Indonesia. Faculty of Medicine, Widya Mandala Catholic University Surabaya, Surabaya, East Java, IndonesiaFaculty of Medicine, Widya Mandala Catholic University Surabaya, East Java, Indonesia Division of Veterinary Anatomy, Department of Veterinary Science, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia; Research Centre for Vaccine Technology and Development, Institute of Tropical Disease, Universitas Airlangga, Surabaya, IndonesiaBackground 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. Conclusion 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. https://univmed.org/ejurnal/index.php/medicina/article/view/1678Wharton’s jellymesenchymal stem cellsisolationflow cytometryregenerative medicine |
| spellingShingle | Yudy Tjahjono Sianty Dewi Bernadette Dian Novita Hendy Wijaya Brilliant Dwi Putra Suryo Kuncorojakti Lucia Hendriati FX Himawan Haryanto Jong Teguh Widodo Franklin Vincentius Malonda Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum Universa Medicina Wharton’s jelly mesenchymal stem cells isolation flow cytometry regenerative medicine |
| title | Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum |
| title_full | Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum |
| title_fullStr | Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum |
| title_full_unstemmed | Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum |
| title_short | Rapid isolation and characterization of Wharton's jelly-derived mesenchymal stem cells maintained in fresh-prepared human AB-serum |
| title_sort | rapid isolation and characterization of wharton s jelly derived mesenchymal stem cells maintained in fresh prepared human ab serum |
| topic | Wharton’s jelly mesenchymal stem cells isolation flow cytometry regenerative medicine |
| url | https://univmed.org/ejurnal/index.php/medicina/article/view/1678 |
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