In situ freeze-fracture of monolayer cell cultures grown on a permeable support.
In situ freeze-fracture of monolayer cell cultures grown on a permeable support.
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在渗透性支持物上生长的单层细胞培养物的原位冷冻断裂。
DOI：
10.1002/jemt.1070220308
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发表时间：
1992
影响因子：
2.5
通讯作者：
Hazen-Martin,D
中科院分区：
文献类型：
--
作者：
Todd,JH;Sens,DA;Sens,MA;Hazen-Martin,D
培养的上皮细胞在可渗透支持物上的生长允许增加细胞分化和评估各种跨细胞和细胞旁转运过程。需要评估这些细胞在相同条件下相应的超微结构特征，促使本实验室开发了一种可靠的方法来生产这些培养物的冷冻断裂复制品。将单元侧朝上的过滤器插入件的部分放置在聚乙烯醇层之间，其中聚酯薄膜条位于聚乙烯醇的上层上。冷冻后，通过从组件中提起聚酯薄膜条使单层破裂。结果是顶端膜的一致断裂足以分析顶端和侧膜域之间的紧密连接密封链、微绒毛分布和膜内颗粒（IMP）分布。该方法利用标准设备和现成材料，最重要的是，允许冷冻断裂和复制未受干扰的细胞单层。© 1992 Wiley利斯公司
The growth of cultured epithelial cells on permeable supports allows increased cell differentiation and the assessment of a variety of transcellular and paracellular transport processes. The need to assess the corresponding ultrastructural characteristics of these cells underidentical conditions prompted this laboratory to develop a reliable method for producing freezefracture replicas of these cultures. Sections of filter inserts with the cell‐side facing up are placed between layers of polyvinyl alcohol with a strip of mylar positioned on the upper layer of polyvinyl alcohol. Following freezing, the monolayer is fractured by lifting the mylar strip from the assembly. The result is a consistent fracture of the apical membrane sufficient for analysis of tight junction sealing strands, microvilli distribution, and intramembranous particle (IMP) distribution between apical and lateral membrane domains. This method utilizes standard equipment and readily available materials and, most importantly, allows the freeze‐fracture and replication of an undisturbed cell monolayer. © 1992 Wiley‐Liss, Inc.