C166 (ATCC® CRL-2581™)

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

Permits Notice: Necessary Permits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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)
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
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 Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin-0.53% (w/v) 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

Subculture Ratio: 1:5 to 1:10
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994
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

Permits Notice: Necessary Permits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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