CAJ | 학술논문

背景：在保留丝素蛋白原有优点的基础上，采用带正电荷的水溶性壳聚糖对其表面进行修饰，可改善细胞在支架材料上的黏附性。目的：验证壳聚糖表面修饰丝素蛋白支架材料与人脂肪间充质干细胞的生物相容性及两者体外构建组织工程脂肪的可行性。方法：将第3代人脂肪间充质干细胞悬液以1×107 L-1浓度接种于壳聚糖表面修饰丝素蛋白支架材料上作为实验组，以单纯的细胞悬液为对照组，MTT法检测细胞在支架材料上的黏附和增殖能力。将第3代人脂肪间充质干细胞悬液以1×109 L-1浓度接种于壳聚糖表面修饰丝素蛋白支架材料上，分别进行成脂诱导培养与高糖培养基常规培养，14 d后行细胞-支架复合物油红O染色与RT-PCR检测。结果与结论：人脂肪间充质干细胞在壳聚糖表面修饰丝素蛋白支架材料上黏附、增殖良好。成脂诱导14 d后，油红 O 染色显示壳聚糖修饰丝素蛋白支架材料上有大量脂肪细胞生成，且过氧化物酶增殖物活化受体γ2基因表达阳性。结果表明壳聚糖表面修饰丝素蛋白支架材料具有良好的体外生物相容性，与人脂肪间充质干细胞共培养可被成功诱导为成熟脂肪细胞。
배경：재보류사소단백원유우점적기출상，채용대정전하적수용성각취당대기표면진행수식，가개선세포재지가재료상적점부성。목적：험증각취당표면수식사소단백지가재료여인지방간충질간세포적생물상용성급량자체외구건조직공정지방적가행성。방법：장제3대인지방간충질간세포현액이1×107 L-1농도접충우각취당표면수식사소단백지가재료상작위실험조，이단순적세포현액위대조조，MTT법검측세포재지가재료상적점부화증식능력。장제3대인지방간충질간세포현액이1×109 L-1농도접충우각취당표면수식사소단백지가재료상，분별진행성지유도배양여고당배양기상규배양，14 d후행세포-지가복합물유홍O염색여RT-PCR검측。결과여결론：인지방간충질간세포재각취당표면수식사소단백지가재료상점부、증식량호。성지유도14 d후，유홍 O 염색현시각취당수식사소단백지가재료상유대량지방세포생성，차과양화물매증식물활화수체γ2기인표체양성。결과표명각취당표면수식사소단백지가재료구유량호적체외생물상용성，여인지방간충질간세포공배양가피성공유도위성숙지방세포。
BACKGROUND:Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve celladhesion on the scaffolds. OBJECTIVE:To verify the biocompatibility of chitosan-modified silk fibroin with human adipose-derived stem cells (hADSCs), and feasibility of constructing tissue engineered adipose in vitro. METHODS:The hADSCs at passage 3 were seeded on chitosan-modified silk fibroin at the concentration of 1×107/L, as the experiment group;at the same cellconcentration, hADSCs were seeded in 96-wel plates as the control group. MTT tests were performed to evaluate the adhesion, growth and proliferation of hADSCs on chitosan-modified silk fibroin. Then hADSCs were implanted on the chitosan-modified silk fibroin scaffolds at the concentration of 1×109/L. The hADSCs seeded onto chitosan-modified silk fibroin complexes were respectively cultured with adipogenic differentiation medium and ordinary high-glucose DMEM. The complexes were stained with oil red O, and detected with RT-PCR after cultured 14 days. RESULTS AND CONCLUSION:The hADSCs adhered to and proliferated on the scaffolds. After cultured with adipogenic differentiation medium for 14 days, oil red O staining demonstrated that there were amount of mature adipocytes on the scaffold. The peroxisome proliferator activated receptorγ2 was positively expressed. The chitosan-modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successful y.