HEL 299 (ATCC® CCL-137)

Organism: Homo sapiens, human  /  Cell Type: fibroblast  /  Tissue: lung  /  Disease: Normal

Organism Homo sapiens, human
Tissue lung
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease Normal
Age fetus
Gender male
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Karyotype normal human male; diploid; stable. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines.
Clinical Data
male
Black
fetus
Receptor Expression
acetylcholine receptor, muscarinic m2 receptor
Virus Susceptibility Vesicular stomatitis, Orsay (Indiana)
Vesicular stomatitis, Glasgow (Indiana)
Human poliovirus 1
Comments
The capacity of this cell line to propagate in culture is limited, and they will eventually senesce. Cells from the current distribution stocks are capable of at least 18 population doublings before the onset of senescence.
M2 muscarinic receptor expression is downregulated following protein kinase C stimulation.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing

Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting
  5. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended

Medium Renewal: Twice per week

Cryopreservation
Culture medium, 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 7,10
D13S317: 11,12
D16S539: 10,11
D5S818: 11,13
D7S820: 8,11
THO1: 7
TPOX: 8,12
vWA: 16
Isoenzymes
G6PD, A
Name of Depositor RG Brackett
References

Peterson WD Jr., et al. Glucose-6-phosphate dehydrogenase isoenzymes in human cell cultures determined by sucrose-agar gel and cellulose acetate zymograms. Proc. Soc. Exp. Biol. Med. 128: 772-776, 1968. PubMed: 5668122

Rousell J, et al. Transcriptional down-regulation of m2 muscarinic receptor gene expression in human embryonic lung (HEL 299) cells by protein kinase C. J. Biol. Chem. 270: 7213-7218, 1995. PubMed: 7706260

Shimamoto T, et al. Inhibition of DLX-7 homeobox gene causes decreased expression of GATA-1 and c-myc genes and apoptosis. Proc. Natl. Acad. Sci. USA 94: 3245-3249, 1997. PubMed: 9096378

Campbell S, et al. Evidence for ethnic tropism of Helicobacter pylori. Infect. Immun. 65: 3708-3712, 1997. PubMed: 9284141

Muralidhar S, et al. Human cytomegalovirus mtrII oncoprotein binds to p53 and down-regulates p53-activated transcription. J. Virol. 70: 8691-8700, 1996. PubMed: 8970996

Basic Documentation
References

Peterson WD Jr., et al. Glucose-6-phosphate dehydrogenase isoenzymes in human cell cultures determined by sucrose-agar gel and cellulose acetate zymograms. Proc. Soc. Exp. Biol. Med. 128: 772-776, 1968. PubMed: 5668122

Rousell J, et al. Transcriptional down-regulation of m2 muscarinic receptor gene expression in human embryonic lung (HEL 299) cells by protein kinase C. J. Biol. Chem. 270: 7213-7218, 1995. PubMed: 7706260

Shimamoto T, et al. Inhibition of DLX-7 homeobox gene causes decreased expression of GATA-1 and c-myc genes and apoptosis. Proc. Natl. Acad. Sci. USA 94: 3245-3249, 1997. PubMed: 9096378

Campbell S, et al. Evidence for ethnic tropism of Helicobacter pylori. Infect. Immun. 65: 3708-3712, 1997. PubMed: 9284141

Muralidhar S, et al. Human cytomegalovirus mtrII oncoprotein binds to p53 and down-regulates p53-activated transcription. J. Virol. 70: 8691-8700, 1996. PubMed: 8970996