To investigate if myxoma cells can be cultured, tissues were cut into 1 to 2 mm piece, washed with Hanks' balanced salt solution (HBSS) (Invitrogen, Carlsbad, CA), and incubated with 0.1% collagenase II for 30 minutes at 37°C with frequent shaking [8 (link)]. Cells were then filtered through 100 μm mesh. The obtained cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) supplemented with 10% fetal bovine serum (FBS) (Hyclone, Logan, UT), 0.1 mM nonessential amino acids, 100 U/mL Penicillin G, 100 μg/mL streptomycin, 2 mmol/L glutamine, and 0.1 mmol/L β-mercaptoethanol [8 (link)]. After 2 to 3 weeks, a population of phase-bright cells appeared over the adhered fibroblast-like cells. These phase-bright cells were collected by two washes with PBS and one wash with cell dissolution buffer (Gibco, Grand Island, NY) at room temperature under microscope monitoring and subcultured with the same medium [8 (link)]. For cardiac and smooth muscle differentiation, myxoma derived cells were cultured under conditions as previously described [8 (link), 9 (link)].
Cell dissolution buffer
Manufactured by Thermo Fisher Scientific
Cell dissolution buffer is a solution used to lyse or break down cells to extract and analyze their intracellular components, such as proteins, nucleic acids, or organelles. It helps solubilize cell membranes and disrupts cellular structures, enabling the release and isolation of the desired cellular contents for further analysis or processing.
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Lab products found in correlation
2 protocols using cell dissolution buffer
1
Myxoma Cell Culture Protocol
2
Isolation and Characterization of Cardiac Stem Cells
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CSCs were isolated from BMT mice as described6 (link)16 (link)17 (link)18 (link)19 (link). In brief, myocardial tissue was cut into 1- to 2-mm piece, washed with Hanks’ balanced salt solution (HBSS) (Invitrogen, Carlsbad, CA), and incubated with 0.1% collagenase II for 30 minutes at 37 °C with frequent shaking. Cells were then filtered through 100-μm mesh. The cardiac cell samples were either directly evaluated for the expression of GFP by FACS or subcultured as described6 (link)16 (link). After 2 to 3 weeks, a population of phase-bright cells appeared over the adhered fibroblast-like cells. These phase-bright cells were collected by two washes with PBS, and one wash with cell dissolution buffer (Gibco, Grand Island, NY) at room temperature under microscope monitoring, GFP+ cells were sorted by FACS, and sub-cultured in poly-lysine coated plates (BD Biosciences) with the same medium. An outline of the experiments is depicted in Fig. 1 .
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