MPRO Cell Line, Clone 2.1 (ATCC® CRL-11422)

Organism: Mus musculus, mouse  /  Tissue: bone marrow  / 

Organism Mus musculus, mouse
Tissue bone marrow
Product Format frozen
Culture Properties suspension
Biosafety Level 2 [Cells contain SV-40 viral DNA sequences]
Gender male
Strain BDF1
Storage Conditions liquid nitrogen vapor phase
Derivation Bone marrow cells of male BDF1 mice were infected with the retroviral vector LRARalpha403SN. The infected cells were subsequently selected in a medium containing G418 and GM-CSF. The infected cells proliferated and stayed immature in morphology.
Clinical Data
Bone marrow cells of male BDF1 mice were infected with the retroviral vector LRARalpha403SN.
male
Comments
RARalpha403 was inserted into the retroviral vector LXSN to generate the recombinant vector designated LRARalpha403SN.
RARalpha403 is a truncated cDNA that encodes a peptide of 403 amino acids containing the N-terminus and the DNA-binding domain.
RARalphacDNA contains a truncation of the sequences coding for the C-terminal 59 amino acids as well as a portion of the 3' untranslated region.
This GM-CSF dependent neutrophil progenitor cell line is capable of differentiating into neutrophils but not monocytes, mast cells or basophils.
The cells exhibit promyelocyte characteristics including the presence of numerous azurophilic primary granules and cell surface epression of mouse neutrophIL-specific antigen 7/4.
They exhibited positive staining for chloroacetate esterase.
Northern blot analysis indicates that the MPRO cells express high levels of retroviral mRNA harboring the truncated RARalpha403 sequence.
Complete Growth Medium Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate containing 10 ng/ml murine granulocyte macrophage colony stimulating factor, 80%; heat-inactivated horse serum, 20%
Subculturing

Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension in fresh medium at 5 x 105 viable cells/mL. Maintain cultures at cell concentrations between 5 x105  and 2 x106 viable cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)

Cryopreservation
Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Name of Depositor Fred Hutchinson Cancer Res. Cntr.
References

Weiler SR, et al. D3: a gene induced during myeloid cell differentiation of Linlo c-Kit+ Sca-1(+) progenitor cells. Blood 93: 527-536, 1999. PubMed: 9885214

Tsai S, Collins SJ. Creating novel hematopoietic cell lines by expressing altered retinoic acid receptors. US Patent 5,830,760 dated Nov 3 1998

Scott LM, et al. E3, a hematopoietic-specific transcript directly regulated by the retinoic acid receptor alpha. Blood 88: 2517-2530, 1996. PubMed: 8839844

Johnson BS, et al. Retinoid X receptor (RXR) agonist-induced activation of dominant-negative RXR-retinoic acid receptor alpha403 heterodimers is developmentally regulated during myeloid differentiation. Mol. Cell. Biol. 19: 3372-3382, 1999. PubMed: 10207061

Du Y, et al. Identification of an interleukin-3-regulated aldoketo reductase gene in myeloid cells which may function in autocrine regulation of myelopoiesis. J. Biol. Chem. 275: 6724-6732, 2000. PubMed: 10702227

Lawson ND, et al. Normal neutrophil differentiation and secondary granule gene expression in the EML and MPRO cell lines. Exp. Hematol. 26: 1178-1185, 1998. PubMed: 9808058

Lawson ND, et al. Isolation and characterization of the cDNA for mouse neutrophil collagenase: demonstration of shared negative regulatory pathways for neutrophil secondary granule protein gene expression. Blood 91: 2517-2524, 1998. PubMed: 9516153

Basic Documentation
References

Weiler SR, et al. D3: a gene induced during myeloid cell differentiation of Linlo c-Kit+ Sca-1(+) progenitor cells. Blood 93: 527-536, 1999. PubMed: 9885214

Tsai S, Collins SJ. Creating novel hematopoietic cell lines by expressing altered retinoic acid receptors. US Patent 5,830,760 dated Nov 3 1998

Scott LM, et al. E3, a hematopoietic-specific transcript directly regulated by the retinoic acid receptor alpha. Blood 88: 2517-2530, 1996. PubMed: 8839844

Johnson BS, et al. Retinoid X receptor (RXR) agonist-induced activation of dominant-negative RXR-retinoic acid receptor alpha403 heterodimers is developmentally regulated during myeloid differentiation. Mol. Cell. Biol. 19: 3372-3382, 1999. PubMed: 10207061

Du Y, et al. Identification of an interleukin-3-regulated aldoketo reductase gene in myeloid cells which may function in autocrine regulation of myelopoiesis. J. Biol. Chem. 275: 6724-6732, 2000. PubMed: 10702227

Lawson ND, et al. Normal neutrophil differentiation and secondary granule gene expression in the EML and MPRO cell lines. Exp. Hematol. 26: 1178-1185, 1998. PubMed: 9808058

Lawson ND, et al. Isolation and characterization of the cDNA for mouse neutrophil collagenase: demonstration of shared negative regulatory pathways for neutrophil secondary granule protein gene expression. Blood 91: 2517-2524, 1998. PubMed: 9516153