Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells
<b>Background/Objectives:</b> Mesenchymal stem cells (MSCs) are regarded as a promising cell type with significant therapeutic benefits owing to their ease of isolation, maintenance, and characterisation. However, repeated passages during cultural maintenance frequently result in cellula...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-03-01
|
| Series: | Pharmaceuticals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1424-8247/18/4/483 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850144800618053632 |
|---|---|
| author | Kavitha Govarthanan Raja Sundari Meenakshi Sundaram Arthi Sunil Richard Siva Chander Chabathula Secunda Rupert Jeswanth Sathyanesan Rama Shanker Verma Naveen Jeyaraman Madhan Jeyaraman Ramya Lakshmi Rajendran Prakash Gangadaran Byeong-Cheol Ahn |
| author_facet | Kavitha Govarthanan Raja Sundari Meenakshi Sundaram Arthi Sunil Richard Siva Chander Chabathula Secunda Rupert Jeswanth Sathyanesan Rama Shanker Verma Naveen Jeyaraman Madhan Jeyaraman Ramya Lakshmi Rajendran Prakash Gangadaran Byeong-Cheol Ahn |
| author_sort | Kavitha Govarthanan |
| collection | DOAJ |
| description | <b>Background/Objectives:</b> Mesenchymal stem cells (MSCs) are regarded as a promising cell type with significant therapeutic benefits owing to their ease of isolation, maintenance, and characterisation. However, repeated passages during cultural maintenance frequently result in cellular senescence, limiting their utility in regenerative medicine. <b>Methods:</b> We investigated the differentiation capability between early- (P3) and late-passage MSCs (>P15) and tested the potential of Wnt agonist 99021 to reverse MSCs using standard cell culture protocols that define minimal criteria for MSCs, primarily tri-lineage differentiation assays, biochemical staining gene expression analysis, and senescence assays. <b>Results:</b> We initially noticed distinct signs of morphological aging between early- (P3) and late-passage MSCs (>P15) and further examined the differentiation capability between early- (P3) and late-passage MSCs (>P15). We found a diminished differentiation potential in late-passage MSCs. Our senescence assay also revealed >P15 cells were able to absorb the senescence dye, indicating that >P15 MSCs underwent senescence. We further demonstrated that CHIR 99021 reversed the differentiation inhibitory potential-mediated impasse of late-passage MSCs by employing tri-lineage specific differentiation assays, biochemical labelling, and gene expression analysis. Senescence assays after CHIR 99021 treatment also revealed no senescence dye uptake at all. <b>Conclusions:</b> Our findings demonstrated that CHIR 99021 Wnt agonist maybe aids in the reversal of MSC aging-related differentiation inhibition glitches and offers a proven demonstrated protocol for rejuvenating late-passage MSCs. Thus, CHIR99021 treatment inherently reverts the tri-lineage potency in late-passage MSCs, and this method could be further employed to ensure a plentiful MSC source for clinical purposes. |
| format | Article |
| id | doaj-art-790fd6297feb47c5ac1d9d7ef2ed31c0 |
| institution | OA Journals |
| issn | 1424-8247 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj-art-790fd6297feb47c5ac1d9d7ef2ed31c02025-08-20T02:28:15ZengMDPI AGPharmaceuticals1424-82472025-03-0118448310.3390/ph18040483Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem CellsKavitha Govarthanan0Raja Sundari Meenakshi Sundaram1Arthi Sunil Richard2Siva Chander Chabathula3Secunda Rupert4Jeswanth Sathyanesan5Rama Shanker Verma6Naveen Jeyaraman7Madhan Jeyaraman8Ramya Lakshmi Rajendran9Prakash Gangadaran10Byeong-Cheol Ahn11Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, Tamil Nadu, IndiaDepartment of Regenerative Medicine and Research, Government Stanley Hospital, Chennai 600001, Tamil Nadu, IndiaDepartment of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, Tamil Nadu, IndiaDepartment of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, Tamil Nadu, IndiaDepartment of Regenerative Medicine and Research, Government Stanley Hospital, Chennai 600001, Tamil Nadu, IndiaDepartment of Regenerative Medicine and Research, Government Stanley Hospital, Chennai 600001, Tamil Nadu, IndiaDepartment of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, Tamil Nadu, IndiaDepartment of Orthopaedics, ACS Medical College and Hospital, Dr MGR Educational and Research Institute, Chennai 600077, Tamil Nadu, IndiaDepartment of Orthopaedics, ACS Medical College and Hospital, Dr MGR Educational and Research Institute, Chennai 600077, Tamil Nadu, IndiaBK21 FOUR KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, Department of Biomedical Sciences, School of Medicine, Kyungpook National University, Daegu 41944, Republic of KoreaBK21 FOUR KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, Department of Biomedical Sciences, School of Medicine, Kyungpook National University, Daegu 41944, Republic of KoreaBK21 FOUR KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, Department of Biomedical Sciences, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea<b>Background/Objectives:</b> Mesenchymal stem cells (MSCs) are regarded as a promising cell type with significant therapeutic benefits owing to their ease of isolation, maintenance, and characterisation. However, repeated passages during cultural maintenance frequently result in cellular senescence, limiting their utility in regenerative medicine. <b>Methods:</b> We investigated the differentiation capability between early- (P3) and late-passage MSCs (>P15) and tested the potential of Wnt agonist 99021 to reverse MSCs using standard cell culture protocols that define minimal criteria for MSCs, primarily tri-lineage differentiation assays, biochemical staining gene expression analysis, and senescence assays. <b>Results:</b> We initially noticed distinct signs of morphological aging between early- (P3) and late-passage MSCs (>P15) and further examined the differentiation capability between early- (P3) and late-passage MSCs (>P15). We found a diminished differentiation potential in late-passage MSCs. Our senescence assay also revealed >P15 cells were able to absorb the senescence dye, indicating that >P15 MSCs underwent senescence. We further demonstrated that CHIR 99021 reversed the differentiation inhibitory potential-mediated impasse of late-passage MSCs by employing tri-lineage specific differentiation assays, biochemical labelling, and gene expression analysis. Senescence assays after CHIR 99021 treatment also revealed no senescence dye uptake at all. <b>Conclusions:</b> Our findings demonstrated that CHIR 99021 Wnt agonist maybe aids in the reversal of MSC aging-related differentiation inhibition glitches and offers a proven demonstrated protocol for rejuvenating late-passage MSCs. Thus, CHIR99021 treatment inherently reverts the tri-lineage potency in late-passage MSCs, and this method could be further employed to ensure a plentiful MSC source for clinical purposes.https://www.mdpi.com/1424-8247/18/4/483MSCssenescencerejuvenationCHIR 99021tri-lineagedifferentiation |
| spellingShingle | Kavitha Govarthanan Raja Sundari Meenakshi Sundaram Arthi Sunil Richard Siva Chander Chabathula Secunda Rupert Jeswanth Sathyanesan Rama Shanker Verma Naveen Jeyaraman Madhan Jeyaraman Ramya Lakshmi Rajendran Prakash Gangadaran Byeong-Cheol Ahn Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells Pharmaceuticals MSCs senescence rejuvenation CHIR 99021 tri-lineage differentiation |
| title | Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells |
| title_full | Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells |
| title_fullStr | Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells |
| title_full_unstemmed | Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells |
| title_short | Inhibition of GSK-3β Restores Differentiation Potential of Late-Passage Mesenchymal Stem Cells |
| title_sort | inhibition of gsk 3β restores differentiation potential of late passage mesenchymal stem cells |
| topic | MSCs senescence rejuvenation CHIR 99021 tri-lineage differentiation |
| url | https://www.mdpi.com/1424-8247/18/4/483 |
| work_keys_str_mv | AT kavithagovarthanan inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT rajasundarimeenakshisundaram inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT arthisunilrichard inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT sivachanderchabathula inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT secundarupert inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT jeswanthsathyanesan inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT ramashankerverma inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT naveenjeyaraman inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT madhanjeyaraman inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT ramyalakshmirajendran inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT prakashgangadaran inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells AT byeongcheolahn inhibitionofgsk3brestoresdifferentiationpotentialoflatepassagemesenchymalstemcells |