Porcine adipose derived stem cells: isolation, characterization and multi-differentiation potential
Baocheng Zhao, Bo Zhao, Zhenjun Wang
Abstract
Baocheng Zhao, Bo Zhao, Zhenjun Wang
Abstract
Objective To investigate the isolation, culture, biological characteristics and multi-differentiation of porcine adipose-derived stem cells (ADSCs) in vitro. Methods ADSCs were obtained from back subcutaneous adipose tissue of porcine by collagenase digestion and cultured in vitro. Flow cytometric analysis and cyto-immumofluorescence staining technique were carried out to detect the immunophenotypes of ADSCs. Reverse transcription-polymerase chain reaction (RT-PCR) analysis and specific staining were used to evaluate the multipotential differentiation phenotype of ADSCs. Results The stem cells isolated from subcutaneous adipose tissue were positive for CD44 (90.2%) and CD105 (99.2%), lacking in CD34 (0.3%) and CD45 (1.2%). After culture in mesenchymal stem cell adipogenic, chondrogenic and osteogenic differentiation basal media respectively for 3, 4 and 3 weeks, the cells were positive for oil red O staining (93%) and alizarin bordeaux staining (94%), also the expression of peroxisome proliferator activated receptor (PPAR)-γ, collagen and aggrecan was detected in the differentiated cells. Conclusion The porcine ADSCs can differentiate towards adipocytes, chondrocytes and osteoblasts when culture in certain medium, and ADSCs can likely served as one of the most optimal autogenous cell sources for tissue engineering. Key words: Porcine; Adipose derived stem cells; Immunophenotype; Induce
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Objective To investigate the isolation, culture, biological characteristics and multi-differentiation of porcine adipose-derived stem cells (ADSCs) in vitro. Methods ADSCs were obtained from back subcutaneous adipose tissue of porcine by collagenase digestion and cultured in vitro. Flow cytometric analysis and cyto-immumofluorescence staining technique were carried out to detect the immunophenotypes of ADSCs. Reverse transcription-polymerase chain reaction (RT-PCR) analysis and specific staining were used to evaluate the multipotential differentiation phenotype of ADSCs. Results The stem cells isolated from subcutaneous adipose tissue were positive for CD44 (90.2%) and CD105 (99.2%), lacking in CD34 (0.3%) and CD45 (1.2%). After culture in mesenchymal stem cell adipogenic, chondrogenic and osteogenic differentiation basal media respectively for 3, 4 and 3 weeks, the cells were positive for oil red O staining (93%) and alizarin bordeaux staining (94%), also the expression of peroxisome proliferator activated receptor (PPAR)-γ, collagen and aggrecan was detected in the differentiated cells. Conclusion The porcine ADSCs can differentiate towards adipocytes, chondrocytes and osteoblasts when culture in certain medium, and ADSCs can likely served as one of the most optimal autogenous cell sources for tissue engineering. Key words: Porcine; Adipose derived stem cells; Immunophenotype; Induce
Key concepts: Adipose tissue, Stem cell, CD44, Mesenchymal stem cell, CD34, Adipogenesis, CD90, Cell biology