摘要
Background： Several studies have revealed that adipose-derived mesenchymal stem cells （ADSCs） can be used as seed cells for the treatment of spinal cord injury （SCI）. Chondroitinase ABC （ChABC） decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI, allowing stem cells to penetrate through the scar and promote recovery of nerve function. This study aimed to establish ADSCs that stably express ChABC （ChABC-ADSCs） and evaluate the migratory capability of ChABC-ADSCs in vitro. Methods： ADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion. Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation. After successful culture, ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained. Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method, ChABC expression was verified using Western blotting, and the migration of ChABC-ADSCs was analyzed using the transwell assay. Results： Secondary collagenase digestion increased the isolation efficiency of primary ADSCs. Following transfection using lentivira[ vectors, the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h （P 〈 0.05）. And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group （P 〈 0.05）, Moreover, ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control （P 〈 0.05）. Conclusions： Secondary collagenase digestion can be used to effectively isolate ADSCs. ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC, and ChABC expression significantly enhances the migratory capacity of ADSCs.
Background： Several studies have revealed that adipose-derived mesenchymal stem cells （ADSCs） can be used as seed cells for the treatment of spinal cord injury （SCI）. Chondroitinase ABC （ChABC） decomposes chondroitin sulfate proteoglycans in the glial scar that forms following SCI, allowing stem cells to penetrate through the scar and promote recovery of nerve function. This study aimed to establish ADSCs that stably express ChABC （ChABC-ADSCs） and evaluate the migratory capability of ChABC-ADSCs in vitro. Methods： ADSCs were obtained from Sprague-Dawley rats using secondary collagenase digestion. Their phenotypes were characterized using flow cytometry detection of cell surface antigens and their stem cell properties were confirmed by induction of differentiation. After successful culture, ADSCs were transfected with lentiviral vectors and ChABC-ADSCs were obtained. Proliferation curves of ChABC-ADSCs were determined using the Cell Counting Kit-8 method, ChABC expression was verified using Western blotting, and the migration of ChABC-ADSCs was analyzed using the transwell assay. Results： Secondary collagenase digestion increased the isolation efficiency of primary ADSCs. Following transfection using lentivira[ vectors, the proliferation of ChABC-ADSCs was reduced in comparison with control ADSCs at 48 h （P 〈 0.05）. And the level of ChABC expression in the ChABC-ADSC group was significantly higher than that of the ADSC group （P 〈 0.05）, Moreover, ChABC-ADSC migration in matrigel was significantly enhanced in comparison with the control （P 〈 0.05）. Conclusions： Secondary collagenase digestion can be used to effectively isolate ADSCs. ChABC-ADSCs constructed using lentiviral vector transfection stably express ChABC, and ChABC expression significantly enhances the migratory capacity of ADSCs.
基金
a grant from the National Natural Science Foundation of China
