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.