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RN46A-B14

RN46A-B14

Catalogue No.

12061303

Cell Line Name

RN46A-B14

Cell Line Description

The clonal cell line, RN46A-B14, was isolated following transfection of the gene encoding rat brain-derived neurotrophic factor (BDNF) into RN46A cells (see ECACC Catalogue number 12061302). RN46A-B14 cells synthesize and secrete biologically active BDNF in vitro and synthesize serotonin (5-HT) following partial membrane depolarization. Two weeks following RN46A-B14 cell transplantation into the adult rat cortex and hippocampus, there is a threefold increase in survival of RN46A-B14 cells compared to RN46A cells. The grafted RN46A-B14 cells immunohistochemically stain for BDNF and 5-HT, while RN46A cells transfected with vector only are negative for both BDNF and 5HT. In addition, RN46A-B14 cells attain more morphologically complex phenotypes, indicating enhanced neuronal differentiation. Autocrine secretion of BDNF by RN46A-B14 cells thus potentiates survival and can be used to deliver both BDNF and 5-HT in vivo.

Characteristics

Tissue of Origin

Embryonic rat (day 13) medullary raphe

Morphology

Fibroblast morphology while proliferating and neuronal on differentiation

Disease

None Stated

Culture Conditions

Cell Type

Fibroblast morphology while proliferating and neuronal on differentiation

Subculture Routine

Split subconfluent cultures (70-80%) 1:2 to 1:5 using 0.05% trypsin/EDTA; 5% CO₂; 33°C. Suggested seeding density 2-4x10,000 cells/cm². Doubling time 25hrs.

Culture Medium

DMEM: Ham's F12 (1:1) + 2mM Glutamine + 10% Foetal Bovine Serum (FBS) + 0.25mg/ml Geneticin (G418) + 0.1mg/ml hygromycin. Alternatively CNS medium can be used (see Kawamoto & Barrett 1986).

Growth Mode

Adherent

Additional Info

Depositor

Dr Scott R Whittemore, Kentucky Spinal Cord Injury Research Centre, 511 S. Floyd St., Rm 616A, University of Louisville School of Medicine, Louisville, KY 40202. USA

Country of Origin

United States

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Eaton MJ, Whittemore SR. 1996 Autocrine BDNF secretion enhances the survival and serotonergic differentiation of raphe neuronal precursor cells grafted into the adult rat CNS. Exp Neurol. 140:105-14.PMID: 8690054.

Bibliography

White LA, Eaton MJ, Castro MC, Klose KJ, Globus MY, Shaw G, Whittemore SR. 1994 Distinct regulatory pathways control neurofilament expression and neurotransmitter synthesis in immortalized serotonergic neurons. J Neurosci. 14:6744-53.PMID: 7965075. Kawamoto JC, Barrett JN.1986 Cryopreservation of primary neurons for tissue culture. Brain Res. 384:84-93.PMID: 3791002.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: RN46A-B14 (ECACC 12061303).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.