Stem Cell Phenotyping
Stem Cell Phenotyping - Related Videos
22:06
Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells
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Cited by 7 •
2007
This protocol details the derivation of transplantable hematopoietic stem cells from mouse embryonic stem cells (ESC) and their subsequent injection into lethally irradiated recipient mice. Briefly, ESC are differentiated as embryoid bodies, which are then infected with retroviral HoxB4 and co-cultured with OP9 stromal cells and hematopoietic cytokines.
12:03
Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors
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Cited by 18 •
2012
A method to analyse the distribution of bone marrow hematopoietic progenitors in flow cytometry as well as to efficiently isolate highly purified hematopoietic stem cells (HSCs) is described. The isolation procedure is essentially based on magnetic enrichment of c-Kit+ cells and cell sorting to purify HSCs for cellular and molecular studies.
09:21
Phenotypic Profiling of Human Stem Cell-Derived Midbrain Dopaminergic Neurons
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2023
This protocol describes the cell culturing of human midbrain dopaminergic neurons, followed by immunological staining and the generation of neuronal phenotypic profiles from acquired microscopic high-content images allowing the identification of phenotypic variations due to genetic or chemical modulations.
10:20
Simultaneous Assessment of Kinship, Division Number, and Phenotype via Flow Cytometry for Hematopoietic Stem and Progenitor Cells
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Cited by 1 •
2023
Presented here is a flow cytometry-based technique that allows for simultaneously measuring the number of cell divisions, surface cell phenotype, and cellular kinship. Those properties can be tested statistically using a permutation-based framework.
09:57
Harnessing the DNA Dye-triggered Side Population Phenotype to Detect and Purify Cancer Stem Cells from Biological Samples
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Cited by 4 •
2017
Methods allowing the characterization and isolation of stem cell populations from biological samples are critical for the advance of stem cell-targeted treatments in cancer and beyond. Here, we provide a detailed protocol for cancer stem cell isolation using the dye-triggered side population phenotype.
FAQs
Surface-marker analysis provides molecular evidence that supports classification of stem cell populations. Patterns of marker expression can be evaluated alongside gene and protein profiles, morphology, proliferation, and differentiation responses rather than interpreted in isolation. This combined approach helps researchers distinguish pluripotent, multipotent, and lineage-committed cells more reliably and assess whether a culture matches its intended identity.
Each measurement captures a different aspect of cell behavior. Molecular assays describe marker, gene, and protein profiles, morphological observations reveal cellular appearance, and functional tests examine proliferation or differentiation responses. Combining these readouts reduces reliance on a single indicator and provides a broader assessment of cell identity, current state, and developmental potential.
Culture conditions and biomaterials can influence how stem cells behave, making the surrounding environment an important variable in phenotypic analysis. In bioengineering, measurements can reveal whether a culture system, material, organoid, or tissue-engineered construct is associated with the intended cellular characteristics. These observations help researchers evaluate and refine engineered environments.
The distinction depends on evaluating multiple characteristics, especially molecular profiles and differentiation responses. Pluripotent, multipotent, and lineage-committed populations can show different patterns of marker expression and developmental behavior. Phenotyping therefore supports classification by examining both what cells express and how they respond when researchers assess their capacity for differentiation.
A workflow begins by selecting measurements relevant to the population and the research question, then collecting molecular, morphological, and functional data. Researchers can examine surface markers, gene and protein profiles, morphology, proliferation, and differentiation responses before integrating the results. The combined findings help determine cell identity, state, developmental potential, and culture quality.
Useful quality-control measurements include surface-marker expression, gene and protein profiles, cell morphology, proliferation, and differentiation responses. Together, these readouts show whether cells retain the expected characteristics within a culture system or engineered construct. Applying the same phenotypic criteria across samples also supports assessment of consistency and reproducibility during development or manufacturing.
In organoid and tissue-engineering studies, phenotyping helps determine how designed environments influence cell behavior and whether resulting populations have the intended characteristics. Researchers can use the information to guide culture-system and biomaterial design, evaluate engineered constructs, and support quality control. The same analyses contribute to reproducible development of bioengineered models.
Phenotypic analysis provides evidence that cells used in disease models or drug-testing systems have the relevant identity and state. It also helps evaluate populations intended for regenerative therapies by examining their characteristics and developmental potential. In each setting, systematic measurements improve interpretation of experimental outcomes and support more reproducible cell-based research.