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.
HCN-2 cells can be induced to differentiate when cultured with a mixture of nerve growth factor (NGF), dibutyryl cyclic adenosine monophosphate (cAMP) and 1-isobutyl-3-methylxanthine (IBMX).
Differentiation is accompanied by mature morphology and slowing of growth (doubling time greater than 120 hours). The growth rate of HCN-2 cells is stimulated by treatment with phorbol esters. A similar line (HCN-1A) is available as ATCC CRL-10442.
The cells stain positively for a number of neuronal markers including neurofilament protein, neuron specific enolase (NSE).
They are also positive for tubulin, vimentin, somatostatin (SST), glutamate, gamma aminobutyric acid (GABA), cholecystokinin - 8 (CCK-8) and vasoactive intestinal peptide (VIP).
NOTE: This cell line is extremely slow growing. May not be able to subculture for 12 to 14 days after recovery. Cultures do not reach 100% confluence (about 80-90%). A change in the fetal bovine serum may help increase the growth rate. Subculture at no higher than a 1:3 ratio every 10 to 12 days when cultures are about 90% confluent.
Ronnett GV, et al. Human neuronal cell line. US Patent 5,196,315 dated Mar 23 1993
