弗式不完全佐剂诱导小鼠腹腔淋巴管瘤模型方法的改进

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目的改进弗式不完全佐剂诱导小鼠腹腔淋巴管瘤模型的建立方法。方法 20只C57BL/6小鼠中的16只按照小鼠体重随机分为两组,分别采取d1、d15(实验Ⅰ组)和d1、d8、d15(实验Ⅱ组)模式每次腹腔注射弗氏不完全佐剂与PBS以1:1体积比配制的乳悬液0.2ml。首次注射后约3个月,脱颈处死小鼠,观察小鼠腹腔内诱导形成的白色肿瘤样组织的生长部位,用游标卡尺记录大小,随后对其进行病理组织学观察,并通过免疫组化表达淋巴管内皮细胞透明质酸受体1(LYVE-1)来进一步证实为淋巴管瘤。另外4只小鼠分别作为两组的对照组(对照Ⅰ组和对照Ⅱ组)按照与实验组对应的注射模式腹腔注射0.2 ml PBS缓冲液。结果实验Ⅰ组、实验Ⅱ组小鼠腹腔内成瘤率均为100%。实验Ⅰ组每只小鼠腹腔成瘤总体积(227.82±124.87)mm3,明显小于实验Ⅱ组的(365.38±78.74)mm3(P<0.05)。实验Ⅰ组、实验Ⅱ组的肿瘤样组织均经HE染色后光学显微镜下观察,及免疫组化均强阳性的表达LYVE-1,证实为淋巴管瘤。对照Ⅰ组、对照Ⅱ组4只小鼠腹腔均未见任何新生物生成。结论利用C57BL/6小鼠腹腔注射弗氏不完全佐剂,以d1、d8、d15注射模式能够诱导获得更多的小鼠腹腔淋巴管瘤。 Objective To improve the method of establishing an intraperitoneal lymphangioma model induced by incomplete Freund’s adjuvant. Methods Sixteen of 20 C57BL / 6 mice were randomly divided into two groups according to the body weight of mice: d1, d15 (experiment Ⅰ) and d1, d8 and d15 (experiment Ⅱ) Incomplete adjuvant and PBS in a volume ratio of 1: 1 formulated suspension 0.2ml. About 3 months after the first injection, the mice were sacrificed and the white tumor-like tissues induced by intraperitoneal injection were observed. The size of the tumor was recorded by vernier calipers, and then observed histopathologically by immunohistochemistry Lymphangioma was further confirmed by lymphatic endothelial cell hyaluronan receptor 1 (LYVE-1). The other 4 mice were used as control group (control group I and control group II) as the control group, and intraperitoneal injection of 0.2 ml PBS buffer according to the injection mode corresponding to the experimental group. Results The tumor formation rate of mice in experimental group I and group II was 100%. The total volume of peritoneal tumor formation in experimental group I was (227.82 ± 124.87) mm3, which was significantly lower than that in experimental group (365.38 ± 78.74) mm3 (P <0.05). Tumor-like tissues of experimental group I and II were all observed by light microscopy after HE staining and strong positive expression of LYVE-1 by immunohistochemistry was confirmed as lymphangioma. Control Ⅰ group, control Ⅱ group 4 mice did not see any neoplasia in the abdominal cavity. Conclusion C57BL / 6 mice were given intraperitoneal Freund’s incomplete adjuvant to induce more intraperitoneal lymphangioma in mice with d1, d8 and d15 injection.
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