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《Fudan University Journal of Medical Sciences》 2014-05
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Biocompatibility of rabbit adipose-derived stem cells (ADSCs) with porous polylactic-co-glycolic acid (PLGA) and chitosan scaffold (CS)

LI Chun-bo;WANG Hong;CHEN Zeng-gan;CHEN Tong-yi;ZHANG Feng;ZHOU Jian-ping;CUI Lei;YIN Jing-bo;Department of Orthopedics,Zhongshan Hospital,Fudan University;Department of General Surgery,Zhongshan Hospital,Fudan University;Department of Plastics,Shanghai Ninth People′s Hospital,School of Medicine,Shanghai Jiaotong University;Department of Polymer Material,School of Materials Science and Engineering,Shanghai University;  
Objective To investigate the biocompatibility of rabbit adipose-derived stem cells(ADSCs)and polylactic-co-glycolic acid(PLGA)/chitosan(CS)scaffold and to provide experiment basis for building tissue-engineered adipose tissue. Methods ADSCs were isolated from subcutaneous adipose tissue in groin area of New Zealand white rabbit with the method of enzymatic digestion.The cells were cultured and passaged to 3-5 generation and its differentiation ability was evaluated.After 3-5 generation ADSCs were harvested,re-suspended with a density of 1×107/mL and replanted into PLGA/CS scaffold,scanning electron microscope was used to reveal the microstructure of PLGA/CS and the growth of ADSCs on PLGA/CS scaffold 1 and 7 days after seeding for the evaluation of biocompatibility between ADSCs and PLGA/CS scaffold.The distribution of ADSCs on PLGA/CS scaffold appended with adipogenic inducing medium or basic culture medium was evaluated with Dil fluorescence and confocal microscopy.The proliferation of ADSCs on PLGA/CS scaffold appended with adipogenic inducing medium or basic culture medium was evaluated with hochest33258.At 1 and7 days post-seeding,the toxicity of PLGA/CS scaffold to ADSCs was assessed by Annexin V/PI influorescence fluid.ADSCs and ADSCs-PLGA/CS complex were cultivated in adipogenic inducing medium for 7 days and the differentiated cells were identified by Nile red stanning. Results Primarily cultured rabbit ADSCs presented fibroblast-like appearance and ADSCs cultured in adipogenic and ostogenic medium could differentiate into adipose cell and bone cells.The proliferation of cells on PLGA/CS scaffold with adipogenic inducing medium or basic culture medium went plateau phase on the 8 day.Scanning electron microscopy(SEM)showed that ADSCs could grow well on the PLGA/CS scaffold and abundant extracelluar matrix(ECM)both on the surface and interior pore of scaffold.Annexin V/PI influorescence staining and confocal microscopy demonstrated that the PLGA/CS scaffold could not affect the activity of ADSCs post-seeding.An amount of lipid droplet within cytoplasm of cells on the PLGA/CS scaffold was revealed by Nile red staining. Conclusions The porous PLGA/CS scaffold and rabbit ADSCs has excellent biocompatibility,which could be used as a tissue engineered scaffold for building adipose tissue.
【Fund】: 上海市科委重点攻关项目(10411953900)~~
【CateGory Index】: R318.08
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