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Organism	Rattus norvegicus, rat
Tissue	liver
Cell Type	fibroblast
Product Format	frozen
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Strain	Buffalo
Storage Conditions	liquid nitrogen vapor phase

Derivation	BRL 3A was derived from buffalo rat liver cells
Genes Expressed	Somatomedin like multiplication stimulating activity (MSA)
Cellular Products	somatomedin like multiplication stimulating activity (MSA)
Comments	The serum-free conditioned medium from this cell line is a source of MSA. MSA is a family of polypeptides that can partially satisfy the serum requirement of chick embryo fibroblasts.

Complete Growth Medium	The base medium for this cell line is Eagle''''s Minimum Essential Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10% This medium is formulated for use with a 5% CO2 in air atmosphere. ATCC tested fetal bovine serum is available as ATCC® Catalog No. 30-2020 (500 mL) and ATCC® Catalog No. 30-2021 (100 mL).
Subculturing	Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium with 10% FBS added. Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. 24 hours later perform a fluid change with serum free growth medium if desired (Nissley SP, et al.. 1977). Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: 2 times per week Note: For 24 hrs after initiating culture from a thawed ampule or after subculturing, grow the cells in culture medium with 10% fetal bovine serum. If desired, after 24 hrs, the cells may be transferred to serum-free medium. Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation	Complete growth medium supplemented with 10% fetal bovine serum and 5% DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions	Temperature: 37°C Atmosphere: Air, 95%; carbon dioxide (CO2), 5% Ref