Clonal Sphere Formation
Clonal Sphere Formation - Related Videos
02:26
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0 Views •
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This video demonstrates a procedure for co-culturing the astrocytes and neurons to form 3D spheres. These 3D spheres provide a distinct platform for examining synapse formation and neural network integration, thereby serving as a potential model for research in developmental neuroscience, neurodegenerative diseases, and neuropharmacological interventions.
03:09
Sphere Formation Assay: An In Vitro Method to Identify Pancreatic Cancer Stem Cells and Screen Therapies
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10:32
Enumeration of Neural Stem Cells Using Clonal Assays
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Cited by 5 •
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Neural stem cells (NSCs) refer to cells which can self-renew and differentiate into the three neural lineages. Here, we describe a protocol to determine NSC frequency in a given cell population using neurosphere formation and differentiation under clonal conditions.
09:23
Turbulence Sphere Method: Evaluating Wind Tunnel Flow Quality
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2023
Source: Jose Roberto Moreto and Xiaofeng Liu, Department of Aerospace Engineering, San Diego State University, San Diego, CA Wind tunnel tests are useful in the design of vehicles and structures that are subjected to airflow during their use. Wind tunnel data are generated by applying a controlled air flow to a model of the object being studied. The test model usually has a similar geometry but is a smaller scale compared to the full-sized object. To ensure accurate and useful data is collected...
12:06
Isolation of CD133+ Liver Stem Cells for Clonal Expansion
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Cited by 22 •
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Here we describe the isolation of CD133 expressing liver stem cells and cancer stem cells from whole murine liver, a process that requires tissue digestion, cell enrichment, and flow cytometry isolation. We include methods for advanced single cell isolation and clonal expansion.
FAQs
The culture conditions create a functional selection. Low-density seeding reduces the likelihood that neighboring cells account for the same sphere, while a nonadherent environment prevents growth based on attachment. Serum-free medium further defines the setting. Cells that continue dividing under these constraints are therefore enriched for survival, proliferative, and self-renewal behaviors measured by the assay.
Sphere number primarily reports how many seeded cells successfully generated spheres, whereas sphere size reflects the extent of growth within those structures. Secondary-sphere formation adds a serial readout: cells from an initial sphere are tested for their ability to generate new spheres. Considering these measurements together helps distinguish initial sphere production from sustained self-renewal capacity.
Changes after genetic or pharmacological manipulation can be assessed by comparing sphere formation under the same defined culture conditions. Differences in sphere number, size, or secondary-sphere production indicate altered cellular behavior in the assay. This makes the method useful for examining how specific interventions affect proliferative or stem-like properties.
Cells are prepared as individual cells, seeded at low density, and maintained in nonadherent, often serum-free medium. The culture is then monitored for free-floating spheres, after which investigators quantify sphere number and size. To test serial self-renewal, cells from primary spheres are used to assess secondary-sphere formation under the defined conditions.
Because cells within a tumor population may differ, individual-cell seeding can expose variation in sphere generation, growth, or secondary-sphere production. These differences reveal that cells may not share identical proliferative or stem-like behavior. Researchers can therefore use the assay to examine whether a population contains functionally distinct subgroups rather than behaving uniformly in culture.
In stem and progenitor cell research, the assay provides a culture-based way to examine whether cells display differing capacities for sphere generation and continued renewal. It also supports studies of differentiation potential by allowing cellular behavior to be evaluated under defined conditions. These comparisons help relate sphere-forming properties to broader questions about cell state and developmental potential.