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DOI: 10.3791/53638-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This protocol describes the isolation of skin-infiltrating leukocytes using enzymatic digestion and density gradient separation. This method is effective for studying cellular and molecular mechanisms in mouse models of skin pathologies, such as atopic dermatitis.
该方案描述了在营养丰富的培养基中对小鼠皮肤进行酶消化,然后进行梯度分离以分离白细胞。 由此衍生的电解槽可用于各种下游应用。这是组织解离机和更苛刻的基于胰蛋白酶和分散酶的组织消化方案的有效、经济且改进的替代方案。
该程序的总体目标是使用酶消化和密度梯度分离分离皮肤浸润的白细胞。该方法可应用于许多皮肤病理(包括特应性皮炎)小鼠模型中的细胞和分子机制的研究。虽然这种方法可以提供对皮肤浸润白细胞生物学的见解,但它也可以应用于其他常驻细胞群的研究。
一般来说,刚接触这种方法的人会努力收集皮肤,其中包括完全去除多余的脂肪和结缔组织。这种方法的视觉演示至关重要,因为皮肤采集和密度梯度处理很难学习,并且需要精确和练习。在治疗后的适当时间点,使用 10 厘米手术剪刀小心地从 8 至 12 周龄的成年雌性 ND4 Swiss-Webster 小鼠身上切除 25 x 25 毫米的剃光侧腹皮肤区域。
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