D10.G4.1 (ATCC® TIB-224)

Organism: Mus musculus, mouse  /  Cell Type: helper T lymphocyte (Th-2)  /  Tissue: lymph node  / 

Organism Mus musculus, mouse
Tissue lymph node
Cell Type helper T lymphocyte (Th-2)
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Age adult
Gender male
Strain AKR/J
Applications
D10.G4.1 has been used extensively to study activation of T cells. These cells can also be used as a reliable and sensitive assay for IL-1 (interleukin-1) response, since D10 cells will proliferate only minimally to con A in the absence of IL-1 or feeder cells.
Storage Conditions liquid nitrogen vapor phase
Clinical Data
male
Antigen Expression
H-2k
Receptor Expression

interleukin 1 (IL-1), expressed; interleukin 1 (IL-1)

Genes Expressed
interleukin 4 (interleukin-4, IL-4); interleukin 5 (interleukin-5, IL-5)
Cellular Products
interleukin 4 (interleukin-4, IL-4); interleukin 5 (interleukin-5, IL-5)
Comments
The cells are specific for egg white conalbumin (Sigma No. C0755) and alloreactive to I-A b, v.
Supernatant media from Con A stimulated D10.G4.1 cells contains IL-4 and IL-5.
Tested and found negative for ectromelia virus (mousepox).
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium:
  • 10% T-STIM with Con A (rat IL-2 culture supplement ) Becton Dickinson, Catalog No. 354115)
  • 10% fetal bovine serum
  • 0.05 mM 2-mercaptoethanol
  • 10 pg/ml mouse IL-1 alpha (R&D Systems, Catalog No. 400ML)

Subculturing
Subculture actively growing suspension cultures before they have reached 1 X 106 cells/mL or the IL-1 and IL-2 will be rapidly depleted and the cells will lose viability. Use inoculation densities of 2 X 105 viable cells/mL.
Medium Renewal: Twice per week
Cryopreservation
Freeze medium: Complete growth medium supplemented with an additional 10% FBS and 5% (v/v) DMSO.
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor C Janeway, J Kaye, P Conrad
References

Kupper T, et al. Growth of an interleukin-2/interleukin 4-dependent T cell line induced by granulocyte-macrophage colony-stimulating factor (GM-CSF). J. Immunol. 138: 4288-4292, 1987. PubMed: 2953804

Kaye J, Janeway CA J. Induction of receptors for interleukin 2 requires T cell Ag:Ia receptor crosslinking and interleukin 1. Lymphokine Res. 3: 175-182, 1984. PubMed: 6436584

Watson J. Continuous proliferation of murine antigen-specific helper T lymphocytes in culture. J. Exp. Med. 150: 1510-1519, 1979. PubMed: 92524

Kaye J, et al. Both a monoclonal antibody and antisera specific for determinants unique to individual cloned helper T cell lines can substitute for antigen and antigen-presenting cells in the activation of T cells. J. Exp. Med. 158: 836-856, 1983. PubMed: 6193236

Kaye J, Janeway CA Jr.. The Fab fragment of a directly activating monoclonal antibody that precipitates a disulfide-linked heterodimer from a helper T cell clone blocks activation by either allogeneic Ia or antigen and self-Ia. J. Exp. Med. 159: 1397-1412, 1984. PubMed: 6232337

Kaye J, et al. Growth of a cloned helper T cell line induced by a monoclonal antibody specific for the antigen receptor: interleukin 1 is required for the expression of receptors for interleukin 2. J. Immunol. 133: 1339-1345, 1984. PubMed: 6235287

Kaye J, Janeway CA Jr.. The alpha- and beta-subunits of a murine T cell antigen/Ia receptor have a molecular weight of 31,000 in the absence of N-linked glycosylation. J. Immunol. 133: 2291-2300, 1984. PubMed: 6237148

Swain SL, et al. Role of antigen in the B cell response. Specific antigen and the lymphokine IL-5 synergize to drive B cell lymphoma proliferation and differentiation to Ig secretion. J. Immunol. 140: 4224-4230, 1988. PubMed: 2453572

Basic Documentation
References

Kupper T, et al. Growth of an interleukin-2/interleukin 4-dependent T cell line induced by granulocyte-macrophage colony-stimulating factor (GM-CSF). J. Immunol. 138: 4288-4292, 1987. PubMed: 2953804

Kaye J, Janeway CA J. Induction of receptors for interleukin 2 requires T cell Ag:Ia receptor crosslinking and interleukin 1. Lymphokine Res. 3: 175-182, 1984. PubMed: 6436584

Watson J. Continuous proliferation of murine antigen-specific helper T lymphocytes in culture. J. Exp. Med. 150: 1510-1519, 1979. PubMed: 92524

Kaye J, et al. Both a monoclonal antibody and antisera specific for determinants unique to individual cloned helper T cell lines can substitute for antigen and antigen-presenting cells in the activation of T cells. J. Exp. Med. 158: 836-856, 1983. PubMed: 6193236

Kaye J, Janeway CA Jr.. The Fab fragment of a directly activating monoclonal antibody that precipitates a disulfide-linked heterodimer from a helper T cell clone blocks activation by either allogeneic Ia or antigen and self-Ia. J. Exp. Med. 159: 1397-1412, 1984. PubMed: 6232337

Kaye J, et al. Growth of a cloned helper T cell line induced by a monoclonal antibody specific for the antigen receptor: interleukin 1 is required for the expression of receptors for interleukin 2. J. Immunol. 133: 1339-1345, 1984. PubMed: 6235287

Kaye J, Janeway CA Jr.. The alpha- and beta-subunits of a murine T cell antigen/Ia receptor have a molecular weight of 31,000 in the absence of N-linked glycosylation. J. Immunol. 133: 2291-2300, 1984. PubMed: 6237148

Swain SL, et al. Role of antigen in the B cell response. Specific antigen and the lymphokine IL-5 synergize to drive B cell lymphoma proliferation and differentiation to Ig secretion. J. Immunol. 140: 4224-4230, 1988. PubMed: 2453572