Hs 895.Sk (ATCC® CRL-7636)

Organism: Homo sapiens, human  /  Cell Type: fibroblast,Melanoma  /  Tissue: skin  /  Disease: normal

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Organism Homo sapiens, human
Tissue skin
Cell Type fibroblast,Melanoma
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease normal
Age 48 years
Gender female
Ethnicity Caucasian
Applications
The two lines provide tumor and normal counterparts for comparative in vitro studies.
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 46; range = 32 to 46
Derivation
This normal skin fibroblast cell line was isolated from the same patient as Hs 895.T (ATCC CRL-7637), a melanoma.
Comments
Part of the NBL Collection. Unlike other cell lines in the NBL Collection, this item has been fully accessioned by ATCC and is covered by the standard warranty.
Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 90%; fetal bovine serum, 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 0.25% (w/v) Trypsin-0.53mM 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.  Add appropriate aliquots of the cell suspension to new culture vessels.
  6.  Incubate cultures at 37°C.

Subcultivation Ratio: 1:2 to 1:4
Medium Renewal: Every 2 to three days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

Cryopreservation Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Notice: Necessary PermitsPermits

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
References

The line was established from apparently normal tissue from a patient who had metastatic melanoma.