CAJ | 학술논문

目的探讨人脂肪干细胞（ADSCs）作为种子细胞复合珊瑚支架材料在体外三维培养下的成骨活性。方法取行脂肪抽吸术患者的脂肪组织，I型胶原酶消化进行培养，贴壁细胞传代，取第2代细胞进行诱导，培养基中添加成骨诱导剂地塞米松、β-甘油磷酸钠、抗坏血酸和维生素D3。成骨诱导后复合珊瑚继续培养，以未诱导的ADSCs复合物为对照，进行生长曲线、DIO染色后的共聚焦荧光显微镜、碱性磷酸酶（AKP）和骨钙蛋白（OCN）生物化学定量检测。结果7～8d后ADSCs在材料上生长进入平台期，诱导ADSCs复合珊瑚7d后生长良好。诱导ADSCs AKP表达随着时间的推移不断增强，而且在检测的各个时间点，诱导ADSCs AKP表达水平均明显高于未诱导ADSCs（均P＜0.05）。诱导ADSCs在珊瑚支架上第7天后检测到OCN表达，并一直增高，且从第7天开始OCN表达均明显高于未诱导ADSCs（均P＜0.05）。结论脂肪干细胞复合珊瑚支架在三维培养下能够向成骨细胞表型分化。
목적탐토인지방간세포（ADSCs）작위충자세포복합산호지가재료재체외삼유배양하적성골활성。방법취행지방추흡술환자적지방조직，I형효원매소화진행배양，첩벽세포전대，취제2대세포진행유도，배양기중첨가성골유도제지새미송、β-감유린산납、항배혈산화유생소D3。성골유도후복합산호계속배양，이미유도적ADSCs복합물위대조，진행생장곡선、DIO염색후적공취초형광현미경、감성린산매（AKP）화골개단백（OCN）생물화학정량검측。결과7～8d후ADSCs재재료상생장진입평태기，유도ADSCs복합산호7d후생장량호。유도ADSCs AKP표체수착시간적추이불단증강，이차재검측적각개시간점，유도ADSCs AKP표체수평균명현고우미유도ADSCs（균P＜0.05）。유도ADSCs재산호지가상제7천후검측도OCN표체，병일직증고，차종제7천개시OCN표체균명현고우미유도ADSCs（균P＜0.05）。결론지방간세포복합산호지가재삼유배양하능구향성골세포표형분화。
Objective To investigate the osteogenic ability of human adipose stem cells (ADSCs) in coral scaffolds in vit-ros. Methods ADSCs were isolated from liposuction aspirates, then digested by type I collagenase;the passage 2 cells were cultured in conditioned medium containing dexamethasone, β- glycerophosphate, ascorbate- 2- phosphate and VD3. The in-duced cells were co- cultured with coral scaffold and non- induced ADSCs- coral construct served as control. The constructs were examined with growth curve, confocal microscopy and quantitative histochemistry. Results The growth curve demonstrat-ed that the proliferation of ADSCs on coral scaffolds reached plateau after 7- 8 d of co- culture. Confocal microscopy and quanti-tative histochemistry showed that ADSCs grew well and began to express osteocalcin on the coral scaffolds after 7d of culture. Quantitative histochemistry demonstrated that the expressions of alkaline phosphatase (AKP) and osteocalcin (OCN) in induced ADSCs- scaffold were higher than those in non- induced ADSCs- scaffold (P<0.05). Conclusion The coral scaffold is a good biomaterial for ADSCs growth and being induced to osteoblasts.