Adipose Derived Stromal Vascular Fraction and Mesenchymal Stem Cells Improve Angiogenesis in a Rat Hindlimb Ischaemia Model

Do Jung Kim; Hyung Min Hahn; Young-Nam Youn; Jun Suk Lee; Il Jae Lee; Sang-Hyun Lim

doi:10.1016/j.ejvs.2023.11.036

Adipose Derived Stromal Vascular Fraction and Mesenchymal Stem Cells Improve Angiogenesis in a Rat Hindlimb Ischaemia Model

Eur J Vasc Endovasc Surg. 2024 May;67(5):828-837. doi: 10.1016/j.ejvs.2023.11.036. Epub 2023 Nov 22.

Authors

Do Jung Kim¹, Hyung Min Hahn², Young-Nam Youn³, Jun Suk Lee², Il Jae Lee², Sang-Hyun Lim⁴

Affiliations

¹ Department of Thoracic and Cardiovascular Surgery, Ajou University Hospital, Ajou University School of Medicine, Suwon, South Korea; Department of Thoracic and Cardiovascular Surgery, Severance Cardiovascular Hospital, Yonsei University College of Medicine, Seoul, South Korea.
² Department of Plastic and Reconstructive Surgery, Ajou University Hospital, Ajou University School of Medicine, Suwon, South Korea.
³ Department of Thoracic and Cardiovascular Surgery, Severance Cardiovascular Hospital, Yonsei University College of Medicine, Seoul, South Korea.
⁴ Department of Thoracic and Cardiovascular Surgery, Ajou University Hospital, Ajou University School of Medicine, Suwon, South Korea. Electronic address: dbricasa@aumc.ac.kr.

PMID: 37995961
DOI: 10.1016/j.ejvs.2023.11.036

Abstract

Objective: This study aimed to investigate the effect of human adipose tissue derived stromal vascular fraction (AD-SVF) and mesenchymal stem cells (AD-MSCs) on blood flow recovery and neovascularisation in a rat hindlimb ischaemia model.

Methods: SVF was isolated using an automated centrifugal system, and AD-MSCs were obtained from adherent cultures of SVF cells. Rats were divided into four groups of six rats each: non-ischaemia (Group 1); saline treated ischaemia (Group 2); SVF treated ischaemia (Group 3); and AD-MSC treated ischaemia (Group 4). Unilateral hindlimb ischaemia was induced in Sprague-Dawley rats via femoral artery ligation. Saline, SVF, or AD-MSCs were injected intramuscularly into the adductor muscle intra-operatively. Cell viability was calculated as the percentage of live cells relative to total cell number. Blood flow improvement, muscle fibre injury, and angiogenic properties were validated using thermal imaging and histological assessment.

Results: The viabilities of SVF and AD-MSCs were 83.3% and 96.7%, respectively. Group 1 exhibited no significant temperature difference between hindlimbs, indicating a lack of blood flow changes. The temperature gradient gradually decreased in SVF and AD-MSC treated rats compared with saline treated rats. In addition, only normal muscle fibres with peripherally located nuclei were observed in Group 1. Groups 3 and 4 exhibited significantly fewer centrally located nuclei, indicating less muscle damage compared with Group 2. Regarding angiogenic properties, CD31 staining of endothelial cells showed similar patterns among all groups, whereas expression of vascular endothelial growth factor, as a crucial angiogenesis factor, was enhanced in the SVF and AD-MSC treated groups.

Conclusion: SVF and AD-MSCs improved blood flow and neovascularisation in a rat hindlimb ischaemia model, suggesting their potential ability to promote angiogenesis. Further extensive research is warranted to explore their potential applications in the treatment of severe lower extremity arterial disease.

Keywords: Adipose derived mesenchymal stem cell; Ischaemia; Neovascularisation; Peripheral arterial disease; Stromal vascular fraction.

MeSH terms

Adipose Tissue* / cytology
Angiogenesis
Animals
Cell Survival
Cells, Cultured
Disease Models, Animal*
Hindlimb* / blood supply
Humans
Injections, Intramuscular
Ischemia* / pathology
Ischemia* / physiopathology
Ischemia* / therapy
Male
Mesenchymal Stem Cell Transplantation* / methods
Mesenchymal Stem Cells
Muscle, Skeletal / blood supply
Neovascularization, Physiologic*
Rats
Rats, Sprague-Dawley*
Regional Blood Flow
Stromal Cells / transplantation