Primary Podocyte Culture
Primary Podocyte Culture - Related Videos
04:19
Deriving Human Kidney Podocytes from Induced Pluripotent Stem Cells: A Procedure for Directed Differentiation of Mature Kidney Podocytes from Stem Cells
0 Views •
2023
In this video, we describe the protocol to derive human kidney podocytes from induced pluripotent stem cells through cell differentiation under defined chemical conditions. The generated podocytes can be used for nephrotoxicity testing and disease modeling.
12:19
Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli
0 Views •
Cited by 25 •
2015
Changes in the intracellular calcium levels in the podocytes are one of the most important means to control the filtration function of glomeruli. Here we explain a high-throughput approach that allows detection of real-time calcium handling and single ion channels activity in the podocytes of the freshly isolated glomeruli.
08:43
Primary Neuronal Cultures
0 Views •
2023
The complexity of the brain often requires neuroscientists to use a simpler system for experimental manipulations and observations. One powerful approach is to generate a primary culture by dissecting nervous system tissue, dissociating it into single cells, and growing those cells in vitro. Primary cultures make neurons and glia easily accessible to the experimental tools required for techniques like genetic manipulation and time-lapse imaging. Furthermore, these cultures represent a highly...
08:14
Using Primary Neurosphere Cultures to Study Primary Cilia
0 Views •
Cited by 3 •
2017
The primary cilium is fundamentally important in neural progenitor cell proliferation, neuronal differentiation, and adult neuronal function. Here, we describe a method to study ciliogenesis and the trafficking of signaling proteins to cilia in neural stem/progenitor cells and differentiated neurons using primary neurosphere cultures.
10:50
Primary Culture of Human Vestibular Schwannomas
0 Views •
Cited by 13 •
2014
Vestibular Schwannomas (VSs) are non-malignant tumors of Schwann cell (SC) origin, associated with mutations in the NF2 tumor suppressor gene. We report a reproducible, efficient protocol for primary human VS cell culture that allows for molecular and cellular experimental manipulation and analysis and recapitulates the heterogeneous nature of human disease.
FAQs
Mechanical and enzymatic dissociation contribute different parts of the isolation process. Mechanical treatment helps disrupt dissected glomerular material, while enzymatic dissociation helps release podocytes from that structure. The sequence is important because the cells must remain viable enough for subsequent attachment and maintenance. Dissociation therefore forms a critical transition between tissue preparation and biochemical analysis.
An extracellular matrix provides the surface needed for podocyte attachment and continued culture. Its presence supports the cellular conditions required for viability and helps maintain a podocyte phenotype rather than merely allowing cells to remain suspended or dispersed. Because attachment and phenotype affect downstream observations, the matrix is an essential part of the culture environment and experimental design.
Cultured podocytes allow researchers to examine podocyte proteins, actin cytoskeleton organization, slit diaphragm biology, and related signaling pathways. These features connect molecular composition with cellular structure and filtration-barrier function. Studying them in the same cellular system can help reveal how biochemical or signaling changes are associated with podocyte behavior and glomerular injury.
A basic workflow begins with dissected glomeruli, followed by mechanical and enzymatic dissociation to release podocytes. The isolated cells are then placed on an extracellular matrix and maintained under defined conditions that support attachment, viability, and podocyte phenotype. Once established, the culture can serve as a controlled system for examining cellular proteins, organization, and responses.
Researchers can use this system when they need a more tractable cellular model for examining podocyte responses under defined conditions. The resulting cultures make it possible to study disease-related stimuli or candidate therapeutics while focusing on podocyte proteins, cytoskeletal organization, slit diaphragm biology, or signaling. This supports controlled investigation of mechanisms that contribute to glomerular injury.
In biochemistry, the culture links molecular measurements with cellular processes relevant to the glomerular filtration barrier. Researchers can investigate protein behavior, actin cytoskeleton organization, slit diaphragm-associated biology, and signaling responses in podocytes. These observations help clarify mechanisms of glomerular injury and provide a cellular context for evaluating responses to disease-related conditions or candidate therapeutics.