Characterization of human adipose-derived stem cells

Abstract

PURPOSE:There is a growing scientific interest in the plasticity and therapeutic potential of adipose-derived stem cells (ASCs), which are multipotent and abundant in adipose tissue and can differentiate in vitro into multiple lineages, including adipocytes, chondrocytes, osteoblasts, neural cells, endothelial cells and cardiomyocytes. The aim of this study was to isolate, cultivate and identify ASCs. METHODS: Human adipose precursor cells were obtained from subcutaneous abdominal tissue. Recently dispersed cells were separated by density centrifugation gradient, cultured and then analyzed. RESULTS: Human ASCs were able to replicate in our culture conditions. The cells maintained their phenotypes throughout the studied period on different passages confirming they suitability for in vitro cultivation. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. Flow cytometry results showed that these cells expressed CD73, CD90 and CD105, (mesenchymal stem-cells markers), contrasting with the lack of expression of CD16, CD34 and CD45 (hematopoietic cells markers). RESUMO OBJETIVO:Há um interesse científico crescente na plasticidade e potencial terapêutico das células-tronco do tecido adiposo humano, células multipotentes e abundantes no tecido adiposo que podem se diferenciar in vitro em múltiplas linhagens celulares, incluindo adipócitos, condrócitos, osteoblastos, células neurais, endoteliais e cardiomiócitos. O objetivo deste estudo foi isolar, cultivar e identificar células-tronco do tecido adiposo humano. MÉTODOS: Células precursoras humanas do tecido adiposo foram obtidas de tecido abdominal subcutâneo. As células recém-dispersas foram separadas por gradiente de centrifugação por densidade, cultivadas e então analisadas. RESULTADOS: As células-tronco do tecido adiposo humano foram capazes de se replicar nas nossas condições de cultivo e mantiveram seu fenótipo em diferentes passagens durante o estudo, confirmando sua adequabilidade para cultivo in vitro. A diferenciação adipogênica, osteogênica e condrogênica também foi induzida, confirmando seu potencial de células-tronco mesenquimais in vitro. Os resultados de citometria de fluxo evidenciaram a expressão dos marcadores de células-tronco mesenquimais CD73, CD90 e CD105, contrastando com a falta de expressão dos marcadores de células hematopoiéticas CD16, CD34 e CD45. CONCLUSÃO: Foi possível isolar células-tronco do tecido adiposo humano por cultivo in vitro sem indução adipogênica, mantendo sua integridade funcional e altos níveis de proliferação. As células demonstraram potencial de diferenciação adipogênico, osteogênico e condrogênico in vitro. Descritores: Adipócitos. Células-Tronco Adultas. Células-Tronco Mesenquimais. Células-Tronco. Tecido Adiposo.Gaiba S et al.

Side-by-Side Comparison of the Biological Characteristics of Human Umbilical Cord and Adipose Tissue-Derived Mesenchymal Stem Cells

“…How to select suitable seed cells as well as their large-scale amplification has been an important issue for tissue engineering [19]. Adipose and umbilical cord tissues are good sources for mesenchymal stem cells, and both these cells have potential for multidifferentiation [20, 21]. In this study, we compared the similarities and differences between ASCs and UC-MSCs, aiming to provide a theoretical basis for clinical selection and application of seed cells.…”

Increased Expression of EGR-1 in Diabetic Human Adipose Tissue-Derived Mesenchymal Stem Cells Reduces Their Wound Healing Capacity

“…The differentiation potential and the expression of mesenchymal stem cell surface markers in nAT-MSCs and dAT-MSCs AT-MSCs are known to possess the ability to differentiate into several types of mesenchymal cells and to express some mesenchymal stem cell markers [26][27][28]. To examine the differentiation potential of dAT-MSCs, we first cultured nAT-MSCs and dAT-MSCs under conditions that would induce differentiation into osteoblasts, adipocytes, and chondrocytes.…”

Revisiting the Advances in Isolation, Characterization and Secretome of Adipose-Derived Stromal/Stem Cells

“…Similarly, cells are also characterized based on expression of their surface markers by using flow cytometry [114]. Characterization of surface markers of ASC is generally carried out by incubating subcultured cells with primary monoclonal and secondary antibodies that are labeled with dyes, such as fluorescein isothiocyanate (FITC), texas red, allophycocyanin (APC), or phycoerythrin (PE) [115,116]. Further, these cells incubated with labeled dye conjugated secondary antibodies are washed.…”