Description Rat Lung Primary Mesenchymal Stem Cells are derived from the lung tissues of day-1 Sprague-Dawley Rats. Rat Lung Primary Mesenchymal Stem Cells are grown in T25 tissue culture flasks pre-coated with gelatin-based solution for 0.5 hour and incubated in Cell Biologics’ Cell Culture Medium for 7-15 days. Cultures are then expanded. Prior to shipping, cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x106 cells per ml and is delivered frozen. Rat Lung Primary Mesenchymal Stem Cells are negative for bacteria, yeast, fungi, and mycoplasma. Cells can be expanded 3-6 passages by the ratio of 1:2 under the cell culture conditions specified by Cell Biologics. Repeated freezing and thawing of cells is not recommended. Mouse Lung Mesenchymal Stem Cells are tested for expression of markers using antibodies, CD29 and CD44 positive by flow cytometry. Suggested Medium M5566 – Mesenchymal Stem Cell Medium /w Kit - 500 ml Laboratory Applications Rat Lung Primary Mesenchymal Stem Cells can be used in assays of standard biochemical procedures performed with cell cultures include RT-PCR, Western blotting, immunoprecipitation, or immunofluorescent flow cytometry. Storage Cell Biologics ships frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen (-180 °C) until ready for experimental use. Live-cell shipment is also available on request. Never can primary cells be kept at -20 °C. Documents Cell-Culture-Protocol-CB General Protocol All cell culture procedures must be conducted in a bio-safety cabinet. Any and all media, supplements, and reagents must be sterilized by filtration through a 0.2 µm filter. Use aseptic technique to prevent microbial contamination. Cryo-preserved cells must be stored in liquid nitrogen or seeded immediately upon arrival. Medium: Review the information provided on the Cell Biologics’ website about appropriate culture media (e.g. serum and other supplements). Use pre-warmed (37°C) cell culture media (30-50 ML) to recover cryo-preserved cells and when changing media or splitting cells. Coating of flasks or dishes: Coat sterile culture dishes or flasks with Gelatin-Based Coating Solution (Cell Biologics, Catalog No. 6950) for 10-30 min and then aspirate the excess solution before seeding cells. Cell recovery from cryovial: • Quickly thaw cells in cryo-vial by incubating them in a 37°C water bath for
