Isolation of human intrahepatic leukocytes for phenotypic and functional characterization by flow cytometry.
Isolation of human intrahepatic leukocytes for phenotypic and functional characterization by flow cytometry.
复制标题
用流式细胞术分离人肝内白细胞进行表型和功能鉴定。
DOI：
10.1016/j.xpro.2022.101356
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发表时间：
2022-06-17
期刊：
影响因子：
--
通讯作者：
Pallett, Laura J.
中科院分区：
文献类型：
--
作者：
Kucykowicz, Stephanie;Amin, Oliver E.;Burton, Alice R.;Swadling, Leo;Schmidt, Nathalie M.;Zakeri, Nekisa;Davies, Jessica;Aidoo-Micah, Gloryanne;Stegmann, Kerstin A.;Easom, Nicholas J.;Jeffery-Smith, Anna;Maini, Mala K.;Pallett, Laura J.
随着人们对组织驻留免疫的日益重视，研究组织特异性免疫细胞对体内平衡和疾病的作用势在必行。在这里，我们提供了一个协议的分离人肝内白细胞（IHL）最大限度地提高活力，纯度和产量。我们的方案可通过组织重量进行缩放，从而允许可再现且有效的IHL释放，其适合于通过流式（或质谱）细胞术进行功能表征、细胞分离和分析。此外，我们提供了一个“指南”，以确定预期的IHL产量每克组织处理。有关本方案使用和执行的完整详细信息，请参见、和。改进的高效和可重复的肝内白细胞分离方案结合酶、机械和密度梯度离心的综合方案流式（质量）细胞术表征单个IHL悬液的详细指南预测每克处理组织中肝内白细胞产量的工具随着对组织固有免疫的日益重视，研究有助于体内平衡和疾病的组织特异性免疫细胞势在必行。在这里，我们提供了一个协议的分离人肝内白细胞（IHL）最大限度地提高活力，纯度和产量。我们的方案可通过组织重量进行缩放，从而允许可再现且有效的IHL释放，其适合于通过流式（或质谱）细胞术进行功能表征、细胞分离和分析。此外，我们提供了一个“指南”，以确定预期的IHL产量每克组织处理。
With the growing appreciation of tissue-resident immunity, studying tissue-specific immune cells contributing to both homeostasis and disease is imperative. Here, we provide a protocol for the isolation of human intrahepatic leukocytes (IHL) maximizing viability, purity, and yield. Our protocol is scalable by tissue weight, allowing for reproducible and efficient IHL liberation suitable for functional characterization, cell isolation, and profiling by flow (or mass) cytometry. Furthermore, we provide a “guide” to determine an expected IHL yield per gram of tissue processed. For complete details on the use and execution of this protocol, please refer to, and. Improved protocol for the efficient and reproducible isolation of intrahepatic leukocytes Comprehensive protocol combining enzymatic, mechanical, and density gradient centrifugation Detailed guide for characterization of a single IHL suspension by flow (mass) cytometry Tool to predict intrahepatic leukocyte yield per gram of tissue processed With the growing appreciation of tissue-resident immunity, studying tissue-specific immune cells contributing to both homeostasis and disease is imperative. Here, we provide a protocol for the isolation of human intrahepatic leukocytes (IHL) maximizing viability, purity, and yield. Our protocol is scalable by tissue weight, allowing for reproducible and efficient IHL liberation suitable for functional characterization, cell isolation, and profiling by flow (or mass) cytometry. Furthermore, we provide a “guide” to determine an expected IHL yield per gram of tissue processed.
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影响因子：
4.6
作者：
Stegmann KA;Robertson F;Hansi N;Gill U;Pallant C;Christophides T;Pallett LJ;Peppa D;Dunn C;Fusai G;Male V;Davidson BR;Kennedy P;Maini MK
通讯作者：
Maini MK
影响因子：
7.3
作者：
Easom NJW;Stegmann KA;Swadling L;Pallett LJ;Burton AR;Odera D;Schmidt N;Huang WC;Fusai G;Davidson B;Maini MK
通讯作者：
Maini MK
影响因子：
16.6
作者：
Zakeri N;Hall A;Swadling L;Pallett LJ;Schmidt NM;Diniz MO;Kucykowicz S;Amin OE;Gander A;Pinzani M;Davidson BR;Quaglia A;Maini MK
通讯作者：
Maini MK
影响因子：
5
作者：
Morsy, MA;Norman, PJ;Vaughan, RW
通讯作者：
Vaughan, RW
DOI：
10.1084/jem.20162115
发表时间：
2017-06-05
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Pallett LJ;Davies J;Colbeck EJ;Robertson F;Hansi N;Easom NJW;Burton AR;Stegmann KA;Schurich A;Swadling L;Gill US;Male V;Luong T;Gander A;Davidson BR;Kennedy PTF;Maini MK
通讯作者：
Maini MK