Neuro-2a (Neuroblastoma, mouse)

Current medium for propagation: Eagle's MEM with non-essential amino acids and Earle's BSS, 90%; FBS, 10%. Clone Neuro-2a was established by R.J. Klebe and F.H. Ruddle (J. Cell Biol. 43: 69A, 1969) from a spontaneous tumor of a strain A albino mouse. This tumor line, designated C1300, was obtained from the Jackson Laboratory, Bar Harbor, Maine. The cells were cloned in F12 medium (Ham), agamma bov. serum, 10%, penicillin and streptomycin and 0.125% agarose. The cloned cells were maintained in F12 (Ham) or minimum essential medium (Eagle) and 10% agamma bov. serum. Neuronal cell types and small round stem cells were observed. Neuro-2a cells produce large quantities of microtubular protein which is believed to play a role in a contractile system which is responsible for axoplasmic flow in nerve cells. The cell line has been used for studies on the mechanism of vinblastine precipitation of microtubular protein, the kinetics of GTP binding to isolated protein, the turnover of microtubules in vivo, and the synthesis and assembly of microtubular protein in vivo and in vitro (Proc. Natl. Acad. Sci. USA 65: 129-136, 1970). A culture in approximately the 150th serial passage was submitted to the American Type Culture Collection in May, 1969. DESCRIPTION OF REPOSITORY REFERENCE SEED STOCK Number of Serial Subcultures from Tissue of Origin: Approximately 167. Freeze Medium: Culture medium, 95%; dimethyl sulfoxide (DMSO), 5%; antibiotic-free. Viability: Approximately 80% (dye exclusion). Culture Medium: Minimum essential medium (Eagle) with non-essential amino acids and Hanks' BSS, 90%; FBS (not inactivated), 10%; antibiotic-free. Growth Characteristics of Thawed Cells: An inoculum of 10(5) viable cells/ml in the above culture medium at 37C, multiplies 15-fold in 7 days. Plating Efficiency: Approximately 1% in the above culture medium. Morphology: Neuron-like and amoeboid-like.

Karyology: Chromosome Frequency Distribution 53 Cells: 2n = 40

Cells: 1 2 1 1 2 1 1 3 1 3 2 4 2 1 4 6 4

Chromosomes: 59-70-75 76-81 82 83 84 85 86 87-89-92 93 94 95 96

2 4 1 1 1 1 1 1 1 1

97 98 99-106-128-174-181-185-190-193

Karyotype unstable within a stemline range of 94 to 98 chromosomes. All the cells contain 6 to 10 large chromosomes with median or submedian centromeres and 2 to 4 minute chromosomes. Sterility: Tests for mycoplasma, bacteria, and fungi were negative. Species: Confirmed as mouse by the cytotoxic-antibody dye-exclusion test. Virus Susceptibility: Susceptible to vesicular stomatitis (Indiana Strain) and herpes simplex viruses. Not susceptible to poliovirus type 1. Reverse Transcriptase: Not detected. Tumorigenicity: Tumors formed in 4/4 strain A albino mice 7 days after subcutaneous injection of 4 X 10(6) cells. Submitted by: R.J. Klebe and F.H. Ruddle, Department of Biology, Yale University, New Haven, CT. Prepared and characterized by: Institute for Medical Research, Camden, NJ, and ATCC, Rockville, MD. Price Code: J