HS-27A (ATCC® CRL-2496)

Organism: Homo sapiens, human  /  Cell Type: human papillomavirus 16 (HPV-16) E6/E7 transformed  /  Tissue: bone marrow/stroma  / 

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Organism Homo sapiens, human
Tissue bone marrow/stroma
Cell Type human papillomavirus 16 (HPV-16) E6/E7 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2
[Cells contain human papilloma viral sequences]
Age 30 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Derivation

Long term bone marrow cells were transformed with the amphotropic retrovirus vector LXSN16E6E7 in the presence of polybrene.

Twenty-seven immortalized clones designed HS-1 to HS-27 were isolated.

Clinical Data
30 years
Caucasian
male
Antigen Expression
vascular cell adhesion molecule 1 (CD106, VCAM-1)
Comments

HS-27A is a subclone of HS-27.

One of these cell lines HS-27A (CRL-2496) has been deposited in the ATCC's general collection. One cell line (HS-5) has been deposited in the Patent Depository (CRL-11882).

HS-27A forms large flattened polygonal shaped cells that exemplify "blanket" cells and maintain numerous intercellular contacts with neighboring cells.

Unlike HS-5, HS-27A secretes low levels of growth factors and does not support proliferation of isolated progenitor cells in cocultures.

HS-27A expresses relatively high levels of vascular cell adhesion molecule 1 (VCAM-1).

HS-27A supports the formation of "cobblestone" areas by isolated cells positive for CD34 but low for CD38.
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: 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 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

Subculture Ratio: 1:4 to 1:5
Medium Renewal: 2 to 3 times a week.
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
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Name of Depositor B Torok-Storb
References

Roecklein BA, Torok-Storb B. Functionally distinct human marrow stromal cell lines immortalized by transduction with the human papilloma virus E6/E7 genes. Blood 85: 997-1005, 1995. PubMed: 7849321

Torok-Storb B, et al. Human marrow stromal cell lines which sustain hematopoieses. US Patent 5,879,940 dated Mar 9 1999

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
Restrictions

These cells are distributed for research purposes only. The line is available with the following restrictions:1. This cell line was deposited at the ATCC by Dr. B. Torok-Storb.The line is provided for research purposes only and is available under the condition that the material will not be used for commercial purposes or be distributed to third parties. The cells are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied.2. Any proposed commercial use of the these cells, or their products must first be negotiated with the Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, C2M-027M, Seattle, WA 98109. Telephone (206) 667-6304, FAX (206-667-4732).

References

Roecklein BA, Torok-Storb B. Functionally distinct human marrow stromal cell lines immortalized by transduction with the human papilloma virus E6/E7 genes. Blood 85: 997-1005, 1995. PubMed: 7849321

Torok-Storb B, et al. Human marrow stromal cell lines which sustain hematopoieses. US Patent 5,879,940 dated Mar 9 1999