超顺磁氧化铁标记对大鼠脂肪干细胞向肝样细胞诱导分化的影响  

Effects of intracellular magnetic labeling of stem cells with superparamagnetic iron oxide on the differentiation capability into hepatocyte-like cells

作　　者：李晏[1,2] 刘再毅[1] 梁长虹[1] 朱杰宁[1] 李晓红[1] 梁家亮[1] 

机构地区：[1]广东省人民医院,广东省医学科学院,广州510080 [2]广东省东莞市人民医院放射科,523018

出　　处：《国际生物医学工程杂志》2012年第6期337-342,I0004,I0005,共8页International Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目（30970798）

摘　　要：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（25μg／m1）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、21d 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。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 ceils. Methods Adipose- derived 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.

关 键 词：超顺磁氧化铁 脂肪干细胞 肝样细胞 标记 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]