Murine Bmsc Isolation
Murine Bmsc Isolation - Related Videos
08:56
Isolation of Murine Embryonic Hemogenic Endothelial Cells
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Cited by 11 •
2016
Hematopoietic stem and progenitor cells (HSPC) derive from specialized (hemogenic) endothelial cells during development, yet little is known about the process by which some endothelial cells specify to become blood forming. We demonstrate a flow-cytometry based method allowing simultaneous isolation of hemogenic endothelial cells and HSPC from murine embryonic tissues.
08:08
Isolation and Activation of Murine Lymphocytes
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Cited by 56 •
2016
Lymphocytes are the major players in adaptive immune responses. Here, we present a lymphocyte purification protocol to determine the physiological functions of the desired molecules in lymphocyte activation in vitro and in vivo. The described experimental procedures are suitable for comparing functional capacities between control and genetically modified lymphocytes.
08:24
Isolation of Murine Coronary Vascular Smooth Muscle Cells
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Cited by 11 •
2016
The purpose of this protocol is to demonstrate the isolation and culture techniques of murine primary vascular smooth muscle cells (VSMCs) from the coronary circulation. Once VSMCs have been isolated, they can be used for many standard culture techniques.
07:51
Isolation of Leukocytes from the Murine Tissues at the Maternal-Fetal Interface
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Cited by 44 •
2015
Described herein is a protocol to isolate and analyze the infiltrating leukocytes of tissues at the maternal-fetal interface (uterus, decidua, and placenta) of mice. This protocol maintains the integrity of most cell surface markers and yields enough viable cells for downstream applications including flow cytometry analysis.
15:20
Murine Pancreatic Islet Isolation
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Cited by 119 •
2007
FAQs
Attachment to tissue-culture plastic provides a practical enrichment step in murine BMSC isolation. During culture, stromal cells remain associated with the surface, whereas many hematopoietic cells are nonadherent and can be removed during medium changes. This difference helps produce a culture suited to studying stromal behavior and subsequent differentiation rather than analyzing an unseparated marrow suspension.
Medium changes act as a selective cleanup step after marrow is introduced into culture. Cells that attach to the plastic are retained, while many nonadherent hematopoietic cells are removed with the displaced medium. Repeating this separation supports development of a stromal-enriched population, which is important when later experiments examine bone formation, adipogenesis, or cellular signaling.
Dissociation converts the harvested marrow into a cell suspension that can be distributed for culture. This step helps expose individual cells to the tissue-culture surface, allowing attachment behavior to distinguish stromal cells from many nonadherent hematopoietic cells. The resulting culture provides a more workable starting population for expansion and characterization than intact marrow material.
The workflow progresses from harvesting mouse long bones to flushing out the marrow, dissociating the collected material, and placing the suspension into culture. Once stromal cells attach, medium changes remove many nonadherent hematopoietic cells. Subsequent culture expansion and characterization prepare the isolated population for differentiation studies, signaling experiments, or testing responses to drugs and biomaterials.
Expanded cultures can be directed toward osteogenic or adipogenic differentiation, allowing investigators to examine processes related to bone and fat-forming potential. The same cells support studies of bone and cartilage formation, cellular signaling, and responses to drugs or biomaterials. These outcomes connect the isolation method to skeletal biology, regenerative research, and musculoskeletal disease models.
The method links bone marrow sampling with a culture system that enriches for adherent stromal cells while reducing many nonadherent hematopoietic cells. Those cultures provide an accessible model for examining interactions between stromal behavior, skeletal tissue formation, and regeneration. Researchers can therefore use the system to investigate tissue repair, stem cell biology, and disease-relevant cellular responses.