Rat Lung Mesenchymal Stem Cells

Rat Primary Lung Mesenchymal Stem Cells

Catalog No. R2303

Suggested Medium:  M5566 – Mesenchymal Stem Cell Medium /w Kit - 500 ml

Product 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.

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 of Cell Biologics Products

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.

Authorized Uses of Cell Biologics Products

Rat Lung Primary Mesenchymal Stem Cells from Cell Biologics are distributed for research purposes only. Our products are not authorized for human use, for in vitro diagnostic procedures, or for therapeutic procedures. Transfer or resale of any Cell Biologics’ cells or products from the purchaser to other markets, organizations or individuals is prohibited by Cell Biologics without the company’s written consent.  Cell Biologics’ Terms and Conditions must be accepted before submitting an order.

Disclaimer

Although Rat Lung Primary Mesenchymal Stem Cells are isolated from laboratory mice testing pathogen-free, investigators should handle the cells that they receive from Cell Biologics with caution, since no test procedure can completely guarantee the absence of infectious agents.

Warranty and Liability

CELL BIOLOGICS’ guarantee applies only to your purchase of CELL BIOLOGICS' cells with CELL BIOLOGICS’ Media and Coating Solution for appropriate cell culture and cell testing following CELL BIOLOGICS’ online protocols within 35 days from the date of product delivery.

General Protocol for Recovering or Freezing Primary Cells

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 <1 min until there is just a small bit of ice left in the vial.

•        Promptly remove the vial and wipe it down with 70% ethanol.

•        Transfer cells from the vial to a sterile centrifuge tube. Add 8-10 ml of pre-warmed Cell Biologics Cell Culture Medium.

•        Flush the vial with an additional 0.5-1 ml of medium to ensure complete transfer of cells to the centrifuge tube.

•        Centrifuge cells at 200 g for 5 minutes.

•        Aspirate the supernatant and resuspend the cell pellet in 6 ml of Cell Biologics’ Cell Culture Growth Medium.

•        Add resuspended cells into a T75 flask pre-coated with Gelatin-Based Coating Solution (Cell Biologics, Catalog No. 6950).

•        Place the T75 flask in a humidified, 5%-CO₂ incubator at 37°C.

•        Change culture media the following day to remove non-adherent cells and replenish nutrients.

•        Change cell culture medium every day when cells are >70% confluent.

•        Cells should be checked daily under a microscope to verify appropriate cell morphology.

Expansion of cultured primary cells:

•        Remove and discard the cell culture media from the flask.

•        Flush the adherent layer 2 times using a 5 ml sterile pipette with sterile PBS (1X) without calcium and magnesium to dislodge loosely attached cells and remove fraction.

•        Remove and discard the wash solution from the flask.

•        Incubate cells with warm (37°C) 0.05% Trypsin-EDTA solution (Cell Biologics, Catalog No. 6915) for 2-5 minutes. Use 3.0 ml of Trypsin-EDTA solution when collecting cells from T75 flasks, and 2 ml when using T25 flasks. As soon as cells have detached (the flask may require a few firm gentle taps), add 8-10 ml of Cell Biologics’ Cell Culture Medium supplemented with 5-10 % FBS to a T25 or T75 flask (the FBS will neutralize the trypsin).

•        Plate cells in fresh flasks or plates precoated with Gelatin-Based Coating Solution in a humidified, 5%-CO₂ incubator at 37°C.

•        Change culture media the following day to remove non-adherent cells and replenish nutrients.

•        Cells should be checked daily under a microscopy to verify appropriate cell morphology.

•        Change culture medium every 24-48 hours. Please note that the medium should be changed every day when cells are >70% confluent to remove non-adherent cells and replenish nutrients. Pre-wash cells with 1X PBS 1-2 times whenever replacing the medium.

We recommend splitting primary cells at the follow ratio:

•        The recommended split ratio for primary murine cells is 1:2 or 1:3.

Procedure for Freezing Cells

Materials:

•      1X Phosphate Buffered Saline (PBS-1X)

•      0.05% Trypsin-EDTA (1X) solution (Cell Biologics, Catalog No. 6915)

•      Tissue Culture Media

•      Cold Freezing Media (10% DMSO, 20% FBS and 70% culture medium, Cell Biologics, Catalog No. 6916).

•      Labeled Cryovials

•      Confluent cells

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