CAJ | 학술논문

目的研究超顺磁氧化铁(SPIO)标记对大鼠脂肪干细胞(ADSCs)向肝样细胞诱导分化的影响.方法 0.25％Ⅱ型胶原酶消化SD大鼠脂肪组织,获取原代ADSCs.采用多聚赖氨酸(PLL)介导SPIO(25 μg/ml)标记ADSCs,以肝细胞生长因子(HGF)作为主要诱导因子,分成标记-诱导组、未标记-诱导组、标记-未诱导组及未标记-未诱导组,后2组分别作为对照.光学显微镜检测标记细胞内的铁摄取.台盼兰染色评价ADSCs的细胞活力.SPIO标记-诱导组和未标记-诱导组细胞均向肝样细胞诱导分化.分别在诱导前、诱导后7、14、21d糖原染色分析肝样细胞内糖原储存；免疫细胞化学染色和RT-PCR分析肝样细胞内白蛋白(ALB)的表达.结果 ADSCs胞浆内铁标记率为100％.SPIO标记组与未标记组的细胞活力差异无统计学意义(P＞0.05).标记诱导组与未标记诱导组在诱导后14d细胞胞浆糖原染色均为阳性；诱导21d后,2组细胞胞浆内染色阳性的细胞增多.14、21 d ALB mRNA和蛋白表达水平逐渐增强.结论 SPIO标记对大鼠ADSCs的生长及其向肝样细胞诱导分化无明显影响.
목적 연구초순자양화철(SPIO)표기대대서지방간세포(ADSCs)향간양세포유도분화적영향.방법 0.25％Ⅱ형효원매소화SD대서지방조직,획취원대ADSCs.채용다취뢰안산(PLL)개도SPIO(25 μg/ml)표기ADSCs,이간세포생장인자(HGF)작위주요유도인자,분성표기-유도조、미표기-유도조、표기-미유도조급미표기-미유도조,후2조분별작위대조.광학현미경검측표기세포내적철섭취.태반란염색평개ADSCs적세포활력.SPIO표기-유도조화미표기-유도조세포균향간양세포유도분화.분별재유도전、유도후7、14、21d당원염색분석간양세포내당원저존；면역세포화학염색화RT-PCR분석간양세포내백단백(ALB)적표체.결과 ADSCs포장내철표기솔위100％.SPIO표기조여미표기조적세포활력차이무통계학의의(P＞0.05).표기유도조여미표기유도조재유도후14d세포포장당원염색균위양성；유도21d후,2조세포포장내염색양성적세포증다.14、21 d ALB mRNA화단백표체수평축점증강.결론 SPIO표기대대서ADSCs적생장급기향간양세포유도분화무명현영향.
Objective To investigate the effects of intracellular magnetic labeling of stem cells with superparamagnetic iron oxide (SPIO) on the cell differentiation capability into hepatocyte-like cells.Methods Adiposederived stem cells (ADSCs) were obtained from the inguinal fat tissue of Sprague-Dawley rats.ADSCs were labeled with co-incubation of poly-l-lysine (PLL) and SPIO (25 μg/mL).Intracellular iron uptake was analyzed qualitatively with light microscopy.The viability of ADSCs was evaluated with trypan blue staining.SPIO-labeled and unlabled ADSCs were subjected to differentiate into hepatocyte-like cells with hepatocyte growth factor (HGF).Liver marker gene such as albumin (ALB) was analyzed by RT-PCR.The cell viability between the labeled cell group and unlabeled cell group adopted two independent sample t-Test.Results Light microscopy results revealed intracytoplasmic iron uptake and nearly 100％ of cell labeling efficiency.SPIO-labeled ADSCs had an unaltered viability as compared with unlabeled ADSCs (P＞0.05).After induction,glycogen storage within cytoplasm can be found in the two group on 14 d and 21 d,and the cells with positive staining increased on day 21.The two groups both express the ALB on 14 d and 21 d,which expressed higher on 21 d.Conclusion Intracellular magnetic labeling with SPIO did not affect the viability and capability of ADSCs to differentiate into hepatocyte-like cells.