Myofibroblast Isolation
Myofibroblast Isolation - Related Videos
06:59
Isolation of Primary Myofibroblasts from Mouse and Human Colon Tissue
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Cited by 43 •
2013
The myofibroblast is an influential stromal cell of the gastrointestinal tract that regulates important physiologic processes in both normal and disease states. We describe a technique that allows for the isolation of primary myofibroblasts from both mouse and human colon tissue, which can be utilized for in vitro experimentation.
06:02
Isolation of CD 90+ Fibroblast/Myofibroblasts from Human Frozen Gastrointestinal Specimens
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Cited by 17 •
2016
Here, a protocol to isolate and establish primary fibroblast/myofibroblast (MF) cultures from frozen gastric, small intestinal, and colonic tissue-yielding cells with a MF phenotype-is presented. These cells express CD90, α-SMA and vimentin. MFs can be used for a variety of functional assays including enzymatic activity and cytokine production.
11:18
Isolation of Myofibroblasts from Mouse and Human Esophagus
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Cited by 9 •
2015
We present a protocol to generate primary cultures of murine and human esophageal stromal cells with a myofibroblast phenotype. Cultured cells have spindle shaped morphology, express α-SMA and vimentin, and lack epithelial, hematopoietic and endothelial cell surface markers. Characterized stromal cells can be used in functional studies of epithelial-stromal interactions.
10:45
Isolation and Characterization of Adult Cardiac Fibroblasts and Myofibroblasts
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Cited by 20 •
2020
Obtaining a pure population of fibroblasts is crucial to studying their role in wound repair and fibrosis. Described here is a detailed method to isolate fibroblasts and myofibroblasts from uninjured and injured mouse hearts followed by characterization of their purity and functionality by immunofluorescence, RTPCR, fluorescence-assisted cell sorting, and collagen gel contraction.
08:59
Human Dupuytren's Ex Vivo Culture for the Study of Myofibroblasts and Extracellular Matrix Interactions
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Cited by 3 •
2015
Dupuytren’s disease (DD) is a fibroproliferative disease of the palm of the hand. Here, we present a protocol to culture resection specimens from DD in a three-dimensional (3D) culture system. Such short-term culture system allows preservation of the 3D structure and molecular properties of the fibrotic tissue.
FAQs
The starting material determines how the workflow is organized and how comparisons are interpreted. Tissue samples require dissection followed by dissociation, whereas primary cultures provide cells that can be studied under controlled culture conditions. Keeping the source consistent helps researchers compare myofibroblast behavior across tissues or experimental conditions without confusing source-related differences with treatment effects.
These markers support assessment of cell identity after recovery from tissue or culture. Morphology provides a visible cellular feature, while α-smooth muscle actin and vimentin provide molecular evidence relevant to the myofibroblast phenotype. Using these readouts together strengthens interpretation by linking the observed cells to the intended population rather than relying on appearance alone.
Growth factors and matrix signals can regulate activation and contraction in isolated myofibroblasts. This makes the cultured cells useful for testing how changes in their surrounding environment affect extracellular-matrix production and contractile behavior. Comparing controlled signal conditions can therefore reveal mechanisms involved in tissue remodeling and help evaluate potential anti-fibrotic interventions.
A typical workflow begins with tissue dissection when tissue is the source, followed by enzymatic or mechanical dissociation to separate the cellular material. The resulting cells are then placed under selective culture conditions intended to support the desired population. Identity is subsequently assessed using morphology together with α-smooth muscle actin and vimentin measurements.
Selective culture conditions are important because they help obtain and maintain a population suitable for controlled study. Researchers should keep the culture environment consistent when comparing tissues, growth-factor treatments, or matrix-related signals. Such control improves the reliability of measurements of activation, contraction, and extracellular-matrix production, which are central outcomes in remodeling and fibrosis experiments.
The technique is useful when researchers need a controlled model of wound healing, fibrosis, or tissue remodeling. Isolated cells allow investigators to examine how growth factors and matrix signals regulate activation and contraction, then compare responses across tissues or experimental conditions. These results can clarify disease mechanisms and support evaluation of possible anti-fibrotic strategies.