【摘要】 制备了可注射的新型温敏性壳聚糖-甘油磷酸钠-胶原(chitosan-glycerophosphate-collagen,C-GP-Co)水凝胶并检测了其与脂肪间充质干细胞(adipose tissue-derived stemcells,ADSCs)的生物相容性.首先将体积分数2.2%的壳聚糖溶液与质量分数50%的β-甘油磷酸钠溶液按一定比例混合后,将2mg/mL胶原溶液加入上述溶液中混合得到壳聚糖-甘油磷酸钠-胶原水凝胶;随后用扫描电镜观察其微观结构,测定了37℃时的成胶时间并测量水凝胶中提取培养基的渗透压;最后将分离、培养和免疫表型鉴定后的ADSCs接种在C-GP-Co水凝胶中培养,倒置显微镜下观察细胞的生长形态,并用Live/Dead Viability-Cytotoxicity试剂盒染色观察细胞凋亡状况,CCK-8试剂盒测定细胞增殖情况.结果表明C-GP-Co水凝胶37℃的成胶时间为12min,水凝胶内部呈海绵状多孔结构,水凝胶中提取培养基的渗透压为310～330mmol/kg.培养7d后,C-GP-Co水凝胶表面的ADSCs贴壁生长,并显示出良好的细胞活性.上述结果表明制备的C-GP-Co水凝胶具有良好的细胞相容性,可作为ADSCs生长增殖的良好支架.更多还原

【Abstract】 An injectable new-type temperature-sensitive hydrogel consisted of chitosan-glycerophosphate-collagen(C-GP-Co) was prepared and the biocompability for adipose tissue-derived stem cells(ADSCs) was detected.The C-GP-Co hydrogel was prepared as follows:chitosan solution(2.2%,volume fraction) and β-glycerophosphate solution(50%,mass fraction) were mixed at a series of proportion,and then,the collagen solution(2 mg/mL)was added into the previous mixture to get the C-GP-Co hydrogel.The gelation time was measured at 37 ℃,and the osmotic pressure of the media within the hydrogels was measured,and then,the micro structure analysis was carried out by SEM.The isolated,cultured and identified ADSCs were seeded into C-GP-Co hydrogels and cell growth status was observed under inverted microscope.The cell apoptosis was detected using Live/Dead Viability-Cytotoxicity Kit,while the proliferation of ADSCs was determined by CCK-8 kits.The experimental results show that the gelation time is about 12 min at 37 ℃,the osmotic pressure of media in the hydrogel is in the normal range of 310-330 mmol/kg and the inner structure of hydrogel is spongy porous.And later,the biocompability of the C-GP-Co hydrogel was further examined.After 7 days′ culture,cells within C-GP-Co hydrogels display adherent cell morphology and high cell viability.It is demonstrated that the C-GP-Co hydrogel has excellent cell compatibility and is a proper scaffold for ADSCs proliferation.更多还原