In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration

Abstract Background Adipose-derived stromal vascular fraction (AdSVF) and adipose-derived mesenchymal stem cells (AdMSCs) are promising tools in regenerative medicine due to their accessibility and therapeutic potential. This study focuses on the in vitro characterization of rat-derived AdMSCs (r-Ad...

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Main Authors: Shajahan Amitha Banu, Khan Sharun, Merlin Mamachan, K. M. Manjusha, Rony S. Emmanuel, Anisha Tomy, Vikash Chandra, Gaurav Kumar Sharma, Abijit M. Pawde, Swapan Kumar Maiti, Amarpal
Format: Article
Language:English
Published: SpringerOpen 2025-05-01
Series:Journal of Basic and Applied Zoology
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Online Access:https://doi.org/10.1186/s41936-025-00462-5
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author Shajahan Amitha Banu
Khan Sharun
Merlin Mamachan
K. M. Manjusha
Rony S. Emmanuel
Anisha Tomy
Vikash Chandra
Gaurav Kumar Sharma
Abijit M. Pawde
Swapan Kumar Maiti
Amarpal
author_facet Shajahan Amitha Banu
Khan Sharun
Merlin Mamachan
K. M. Manjusha
Rony S. Emmanuel
Anisha Tomy
Vikash Chandra
Gaurav Kumar Sharma
Abijit M. Pawde
Swapan Kumar Maiti
Amarpal
author_sort Shajahan Amitha Banu
collection DOAJ
description Abstract Background Adipose-derived stromal vascular fraction (AdSVF) and adipose-derived mesenchymal stem cells (AdMSCs) are promising tools in regenerative medicine due to their accessibility and therapeutic potential. This study focuses on the in vitro characterization of rat-derived AdMSCs (r-AdMSCs) isolated from AdSVF (r-AdSVF) for mesodermal tissue regeneration. Results r-AdSVF was isolated through enzymatic digestion of subcutaneous ventral abdominal fat depots, yielding an average of 5.39 ± 0.73 × 106 cells per gram of adipose tissue. The r-AdSVF was differentiated into AdMSCs under optimized culture conditions, with P3 cells achieving 90% confluence within five days and exhibiting fibroblast-like morphology. Comprehensive characterization included morphological analysis, immunocytochemistry, gene expression studies, trilineage differentiation (osteogenic, chondrogenic, and adipogenic), and colony-forming unit assays. r-AdMSCs showed positive expression of mesenchymal markers (CD73, CD90, CD105) and negative expression of hematopoietic markers (CD34, CD45), confirming their stem cell identity. Colony-forming assays demonstrated robust osteogenic differentiation potential, supporting their application in bone regeneration. Conclusions This study highlights the properties of r-AdSVF and r-AdMSCs while emphasizing the need for improved characterization and standardization to advance their use in mesodermal tissue engineering. By establishing a foundational framework, this research contributes to developing safe, effective, and accessible regenerative therapies for mesodermal tissue repair and regeneration.
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spelling doaj-art-59e87641adef4bf899404527a56514002025-08-20T02:03:38ZengSpringerOpenJournal of Basic and Applied Zoology2090-990X2025-05-0186111410.1186/s41936-025-00462-5In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regenerationShajahan Amitha Banu0Khan Sharun1Merlin Mamachan2K. M. Manjusha3Rony S. Emmanuel4Anisha Tomy5Vikash Chandra6Gaurav Kumar Sharma7Abijit M. Pawde8Swapan Kumar Maiti9Amarpal10Division of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Physiology and Climatology, ICAR-Indian Veterinary Research InstituteCentre for Animal Disease Research and Diagnosis, ICAR-Indian Veterinary Research InstituteDivision of Physiology and Climatology, ICAR-Indian Veterinary Research InstituteCentre for Animal Disease Research and Diagnosis, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteDivision of Surgery, ICAR-Indian Veterinary Research InstituteAbstract Background Adipose-derived stromal vascular fraction (AdSVF) and adipose-derived mesenchymal stem cells (AdMSCs) are promising tools in regenerative medicine due to their accessibility and therapeutic potential. This study focuses on the in vitro characterization of rat-derived AdMSCs (r-AdMSCs) isolated from AdSVF (r-AdSVF) for mesodermal tissue regeneration. Results r-AdSVF was isolated through enzymatic digestion of subcutaneous ventral abdominal fat depots, yielding an average of 5.39 ± 0.73 × 106 cells per gram of adipose tissue. The r-AdSVF was differentiated into AdMSCs under optimized culture conditions, with P3 cells achieving 90% confluence within five days and exhibiting fibroblast-like morphology. Comprehensive characterization included morphological analysis, immunocytochemistry, gene expression studies, trilineage differentiation (osteogenic, chondrogenic, and adipogenic), and colony-forming unit assays. r-AdMSCs showed positive expression of mesenchymal markers (CD73, CD90, CD105) and negative expression of hematopoietic markers (CD34, CD45), confirming their stem cell identity. Colony-forming assays demonstrated robust osteogenic differentiation potential, supporting their application in bone regeneration. Conclusions This study highlights the properties of r-AdSVF and r-AdMSCs while emphasizing the need for improved characterization and standardization to advance their use in mesodermal tissue engineering. By establishing a foundational framework, this research contributes to developing safe, effective, and accessible regenerative therapies for mesodermal tissue repair and regeneration.https://doi.org/10.1186/s41936-025-00462-5Adult stem cellsRegenerative therapyLipoaspirateAdipose cell therapyBone regenerationLipoaspirate prolotherapy
spellingShingle Shajahan Amitha Banu
Khan Sharun
Merlin Mamachan
K. M. Manjusha
Rony S. Emmanuel
Anisha Tomy
Vikash Chandra
Gaurav Kumar Sharma
Abijit M. Pawde
Swapan Kumar Maiti
Amarpal
In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
Journal of Basic and Applied Zoology
Adult stem cells
Regenerative therapy
Lipoaspirate
Adipose cell therapy
Bone regeneration
Lipoaspirate prolotherapy
title In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
title_full In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
title_fullStr In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
title_full_unstemmed In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
title_short In vitro isolation and characterization of rat adipose-derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
title_sort in vitro isolation and characterization of rat adipose derived stromal vascular fraction and mesenchymal stem cells for mesodermal tissue regeneration
topic Adult stem cells
Regenerative therapy
Lipoaspirate
Adipose cell therapy
Bone regeneration
Lipoaspirate prolotherapy
url https://doi.org/10.1186/s41936-025-00462-5
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