Hepatocyte-specific Pten-deficient mouse

Liver-specific Pten deficiency driven by albumin promoter leads to steatosis that progresses to a NASH-like stage characterized by inflammation and fibrosis before eventually developing liver tumors; tumors were present in 66% of males and 30% of females at 40-44 weeks, with HCC reaching 83% in males and 50% in females at 74-78 weeks.

结局指标

获得可供光学显微镜观察的 H&E 染色组织切片：细胞核蓝紫色、细胞质与结缔组织纤维粉红色，用于观察组织中细胞的整体结构与形态细节。本方案本身不产出病理评分数值，也不提供特定分子标志物的定量结果。

实验动物

Hematoxylin and Eosin (H&E) staining for histopathology

年龄: as reported in cited studies
组织: liver

实验步骤

脱蜡（Dewaxing）石蜡切片置于二甲苯中脱蜡 3 次，每次 00:03:00。复水（Rehydration）石蜡切片依次置于 100% 乙醇中 3 次、每次 00:03:00，再置于 95% 乙醇中 1 次、00:03:00，随后用水漂洗。苏木素核染色（Hematoxylin staining）石蜡切片以苏木素染液染色 00:00:20，随后在流水下冲洗 00:01:00。分化与返蓝（Differentiation and bluing）石蜡切片置于返蓝液中 00:01:00，随后在流水下冲洗 00:01:00；此步使细胞核呈现蓝色。伊红染色（Eosin staining）石蜡切片以伊红复染 00:00:10；此步使细胞质成分与结缔组织纤维呈现粉红色。脱水与透明（Dehydration and Clearing）石蜡切片依次置于 95% 乙醇中 00:02:00、100% 乙醇中 2 次、每次 00:02:00，随后置于二甲苯中 2 次、每次 00:02:00。封片（Mounting）石蜡切片以 Permount 封片介质加盖玻片封固。

模型身份

模型名称: Hepatocyte-specific Pten-deficient mouse
物种: mouse
NCBI Taxonomy ID: 10090
基因型／合子状态: Hepatocyte-specific Pten null (Albumin-Cre; Pten floxed)

生物学特性

表型出现阶段: Steatosis is established early; steatohepatitis-like inflammation and fibrosis emerge around 10 weeks of age; hepatic tumors appear at 40-44 weeks and HCC by 74-78 weeks.
适用研究问题: Used to model the natural progression of NAFLD to NASH and NASH-associated hepatocellular carcinoma in a spontaneous genetic model; evaluates interventions that modify progression from steatohepatitis to HCC.
模型局限: The evidence adopted here for hepatocyte-specific Pten deficiency comes from review recommendations rather than from direct use of the model in the primary studies of this package; hepatocyte-specific Pten deficiency is a purely genetic model and does not reproduce the metabolic etiology of human NAFLD (no obesity or insulin resistance); the original studies used mixed genetic backgrounds and the precise Cre construct details and substrain composition are not reported in the sources adopted here.

实验方法

组织学分析

实验方案

Hematoxylin and Eosin (H&E) staining for histopathology
本方案用于固定、石蜡包埋组织切片（4 μm）的苏木素-伊红（H&E）染色：经脱蜡、复水、苏木素核染、返蓝、伊红复染、脱水透明与封片七个步骤，使细胞核呈蓝紫色、细胞质与结缔组织纤维呈粉红色，供光学显微镜下观察组织中细胞与整体结构形态。

配套试剂与材料

组织学二甲苯（混合异构体）
脱蜡剂
Ethanol
复水梯度溶剂（去除二甲苯）
95% 乙醇溶液
复水梯度溶剂
自来水（组织学染色用）
复水后漂洗用水（后续各流水冲洗共用）
Epredia Signature Series 苏木素染液（7221）
细胞核染色
Epredia Signature Series 返蓝液（7301）
返蓝（使细胞核转蓝）
Epredia Signature Series 伊红 Y 染液（7111）
细胞质与结缔组织纤维复染
Permount 组织学封片剂（Fisher SP15）
封片介质

参考文献

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