IN VITRO INVESTIGATION OF THE EFFECT OF STROMAL VASCULAR FRACTION ISOLATED BY MECHANICAL AND ENZYMATIC METHODS ON WOUND HEALING

Yıl 2020, Cilt: 83 Sayı: 3, 241 - 246, 29.06.2020

Kemal Tunç Tiryaki

https://doi.org/10.26650/IUITFD.2019.0051

Öz

Objective: The role of Stromal Vascular Fraction (SVF) cell suspension obtained from adipose tissue in wound healing is gaining importance day by day. In this study, the role of SVF cell population, isolated by enzymatic and mechanical methods in wound healing, will be examined in vitro. Material and Method: An SVF cell cocktail was isolated by mechanical and enzymatic methods after adipose tissue was obtained from 10 different patients by the liposuction method (the adipose tissue was divided into two equal groups for enzymatic and mechanical methods). After the isolation process is done, cell viability and cell number were counted using flow cytometry, and cell proliferation rates were measured by WST-1 assay. The Wound Healing Scratch Assay test, which is used as an in vitro model for wound healing, was performed with cell cocktails obtained from both groups. After isolation, the level of expression of the collagen type 1 gene, known to be involved in wound healing, was measured in both groups. Results: While the SVF (E-SVF) cell cocktail obtained bythe enzymatic method was 1,52x106/ml (±3,63, n=10, p=0,015), the number of SVF (M-SVF) cell cocktails obtained by mechanical method was 0,67x106/ml (±1,69, n=10, p=0,015). No significant difference was observed between cell viability of SVF cell cocktails isolated by two different methods. The M-SVF cell cocktail had a 10% higher cell proliferation rate compared to E-SVF. Furthermore, M-SVF cell migration rates and Col 1 gene levels were found to be higher than E-SVF cells. Conclusion: Although the cell number of the M-SVF cell cocktail was less than the cell number of the E-SVF cell cocktail, the collagen type 1 gene level of the M-SVF cell cocktail was found to be higher, so the M-SVF cell cocktail had higher wound healing properties.

Anahtar Kelimeler

Stromal vascular fraction, wound healing, stem cell

MEKANİK VE ENZİMATİK YÖNTEM İLE İZOLE EDİLEN STROMAL VASKÜLER FRAKSİYON’UN YARA İYİLEŞMESİ ÜZERİNDEKİ ETKİSİNİN IN VİTRO İNCELENMESİ

Öz

Amaç: Yağ dokusundan elde edilen Stromal Vasküler Fraksiyon (SVF) hücre süspansiyonun yara iyileşmesindeki rolü gün geçtikçe önem kazanmaktadır. Bu çalışmada enzimatik ve mekanik yöntemler ile izole edilen SVF hücre popülasyonun yara iyileşmesindeki rolü incelenecektir. Gereç ve Yöntem: 10 farklı hastadan yağ alma yöntemi ile edilen yağ dokusu (eşit miktarda iki farklı gruba bölündükten sonra) mekanik ve enzimatik yöntemler kullanılarak SVF hücre kokteyli izole edilmiştir. İzolasyon sonrasında hücre canlılığı ve hücre sayısı akış sitometrisi kullanılarak sayılmış, hücre proliferasyon hızları WST-1 testi ile ölçümlenmiştir. Yara iyileşmesinde in-vitro model olarak kullanılan Scratch Assay testi her iki grup ile elde edilen hücre kokteylleri ile için gerçekleştirilmiştir. İzolasyon sonrasında, yara iyileşmesinde rol aldığı bilinen kolajen tip 1 geninin ifade edilme seviyesi her iki grup içinde ölçümlenmiştir. Bulgular: Enzimatik yöntem ile elde edilen SVF (E-SVF) hücre kokteyli 1,52x106/ml (±3,63, n=10, p=0,015) iken, mekanik yöntem ile elde edilen SVF (M-SVF) hücre kokteyli sayısı 0,67x106/ ml (±1,69, n=10, p=0,015) olarak ölçümlenmiştir. İki farklı yöntem ile izole edilen SVF hücre kokteylinin izolasyon sonrası hücre canlıkları arasında anlamlı bir fark gözlenmemiştir. M-SVF hücre kokteylinin, E-SVF ile karşılaştırıldığında %10 daha fazla hücre proliferasyon hızına sahip olduğu görülmüştür. Ayrıca, M-SVF hücrelerinin migrasyon hızlarının ve Col 1 gen seviyelerinin E-SVF hücrelerine oranla daha fazla olduğu saptanmıştır. Sonuç: M-SVF hücre kokteylinin hücre sayısının E-SVF hücre kokteylinin hücre sayısından az olmasına rağmen, M-SVF hücre kokteylinin kolajen tip 1 gen seviyesinin arttığı görülmüş, böylece M-SVF hücre kokteylinin daha yüksek yara iyileşme özelliklerine sahip olduğunu gözlemlenmiştir.

Anahtar Kelimeler

Stromal vasküler fraksiyon, kök hücre, yara iyileşmesi

Kaynakça

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