title: Layer-shaped alginate hydrogels enhance the biological performance of human adipose-derived stem cells.
abstract: BACKGROUND
  The reconstruction of adipose tissue defects is often challenged by the complications that may occur following plastic and reconstructive surgery, including donor-site morbidity, implant migration and foreign body reaction. To overcome these problems, adipose tissue engineering (ATE) using stem cell-based regeneration strategies has been widely explored in the last years. Mounting evidence has shown that adipose-derived stem cells (ADSCs) represent a promising cell source for ATE. In the context of a small number of reports concerning adipose tissue regeneration using three-dimensional (3-D) systems, the present study was designed to evaluate the biological performance of a novel alginate matrix that incorporates human ADSCs (hADSCs).
  
  
  RESULTS
  Culture-expanded cells isolated from the stromal vascular fraction (SVF), corresponding to the third passage which showed the expression of mesenchymal stem cell (MSC) markers, were used in the 3-D culture systems. The latter represented a calcium alginate hydrogel, obtained by the diffusion of calcium gluconate (CGH matrix), and shaped as discoid-thin layer. For comparative purposes, a similar hADSC-laden alginate hydrogel cross-linked with calcium chloride was considered as reference hydrogel (RH matrix). Both hydrogels showed a porous structure under scanning electron microscopy (SEM) and the hADSCs embedded displayed normal spherical morphologies, some of them showing signs of mitosis. More than 85% of the entrapped cells survived throughout the incubation period of 7 days. The percentage of viable cells was significantly higher within CGH matrix at 2 days post-seeding, and approximately similar within both hydrogels after 7 days of culture. Moreover, both alginate-based hydrogels stimulated cell proliferation. The number of hADSC within hydrogels has increased during the incubation period of 7 days and was higher in the case of CGH matrix. Cells grown under adipogenic conditions for 21 days showed that both analyzed 3-D culture systems support adipogenic differentiation in terms of neutral lipid accumulation and perillipin expression. Furthermore, the cells encapsulated in CGH matrix displayed a more differentiated phenotype.
  
  
  CONCLUSIONS
  The results of this study suggest that both CGH and RH matrices successfully support the survival and adipogenesis of hADSC. An enhancement of biological performance was detected in the case of CGH matrix, suggesting its promising application in ATE.
authors: Department of Biochemistry and Molecular Biology, University of Bucharest, 91-95 Splaiul Independentei, sect 5, Bucharest, Romania.
authors: Bianca
authors: B
authors: Galateanu
authors: Doina
authors: D
authors: Dimonie
authors: Eugeniu
authors: E
authors: Vasile
authors: Sorin
authors: S
authors: Nae
authors: Anisoara
authors: A
authors: Cimpean
authors: Marieta
authors: M
authors: Costache
mesh_terms: D050156:Adipogenesis; D000273:Adipose Tissue / Q000166:cytology*; D000464:Alginates / Q000737:chemistry*; D002122:Calcium Chloride / Q000737:chemistry; D002125:Calcium Gluconate / Q000737:chemistry; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D016130:Immunophenotyping; D016062:Porosity; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering
chemical_list: D000464:Alginates; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D002122:Calcium Chloride; D002125:Calcium Gluconate