C166 (ATCC® CRL-2581™)

Organism: Mus musculus, mouse  /  Tissue: yolk sac  /  Cell Type: endothelial

Organism Mus musculus, mouse
Tissue
yolk sac
Cell Type endothelial
Product Format frozen
Morphology endothelial
Culture Properties adherent
Biosafety Level 1
Age 12 day embryo
Applications
The cell line should prove useful for the study of endothelial cell differentiation and for the determination of the mechanisms underlying the establishment of organ-specific endothelial cell heterogeneity.

The cell line can support the stable proliferation of multipotential hematopoietic stem cells, thus generating adequate numbers of cells for study of the mechanisms involved in their subsequent development and differentiation.

 
Storage Conditions Liquid nitrogen vapor phase
Derivation
The C166 cell line was established from cells from F1 embryos obtained by mating a female NMRI/GSF mouse with a male CD-1 mouse that was transgenic for the human fes (fps/fes) proto-oncogene.
Antigen Expression
vascular cell adhesion molecule 1 (CD106, VCAM-1)
RefWang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159
vascular addressin +
RefWang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159
Receptor Expression
acetylated low density lipoprotein (LDP)
RefWang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159
Oncogene fes + (fps/fes +) RefWang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159
Genes Expressed
angiotensin converting enzyme (ACE),fes + (fps/fes +),vascular cell adhesion molecule 1 (CD106, VCAM-1),vascular addressin +
Comments
These mice express multiple copies of an activated allele of the human fes (fps/fes) proto-oncogene and display hypervascularity progressing to multifocal hemangiomas.

C166 cells exhibit normal endothelial characteristics, such as rearrangement into tubelike structures when placed on Matrigel and retention of cobblestone morphology at confluence.
The cells constitutively express the vascular addressin identified by antibody MECA-99.

The cell line expresses high levels of the cytoplasmic protein-tyrosine kinase encoded by the fes (fps/fes) proto-oncogene.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Protocol:
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
    Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:10 is recommended
    Medium Renewal: Every 2 to 3 days
    Cryopreservation
    Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
    Storage Temperature: Liquid nitrogen vapor phase
    Culture Conditions
    Temperature: 37°C
    Name of Depositor R Auerbach
    References

    Wang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159

    Lu LS, et al. In vitro and in vivo differentiation into B cells, T cells, and myeloid cells of primitive yolk sac hematopoietic precursor cells expanded > 100-fold by coculture with a clonal yolk sac endothelial cell line. Proc. Natl. Acad. Sci. USA 93: 14782-14787, 1996. PubMed: 8962132

    Wang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159

    Basic Documentation
    Product Sheet
    Product Sheet
    Certificate of Analysis
    Certificate of Analysis
    MSDS
    MSDS
    References

    Wang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159

    Lu LS, et al. In vitro and in vivo differentiation into B cells, T cells, and myeloid cells of primitive yolk sac hematopoietic precursor cells expanded > 100-fold by coculture with a clonal yolk sac endothelial cell line. Proc. Natl. Acad. Sci. USA 93: 14782-14787, 1996. PubMed: 8962132

    Wang SJ, et al. Isolation and propagation of yolk-sac-derived endothelial cells from a hypervascular transgenic mouse expressing a gain-of-function fps/fes proto-oncogene. In Vitro Cell. Dev. Biol. Anim. 32: 292-299, 1996. PubMed: 8792159