HS-5 (ATCC® CRL-11882)

Organism: Homo sapiens, human  /  Cell Type: HPV-16 E6/E7 transformed  /  Tissue: bone marrow/stroma  /  Disease: normal

Organism Homo sapiens, human
Tissue bone marrow/stroma
Cell Type HPV-16 E6/E7 transformed
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 [Cells contain human papilloma viral sequences]
Disease normal
Age 30 years
Gender male
Ethnicity Caucasian
Applications HS-23 and HS-27A secrete low levels of growth factors and do not support proliferation of isolated progenitor cells in cocultures.

The cell line can also be used as a feeder layer in ex vivo bone marrow cultures or in colony forming assays.

Storage Conditions liquid nitrogen vapor phase
Images
Derivation
Long term bone marrow cells were transformed with the amphotropic retrovirus vector LXSN16E6E7 in the presence of polybrene. Twenty-seven immortalized clones designated HS-1 to HS-27 were isolated.
Clinical Data
30 years
Caucasian
male
Genes Expressed granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-CSF (GM-CSF), macrophage-CSF (M-CSF), Kit ligand (KL), macrophage-inhibitory protein-1 alpha, interleukin-1 alpha (IL-1alpha), IL-1beta, IL-1RA, IL-6, IL-8, IL-11, and leukemia inhibitory factor (LIF)
Comments

One of these cell lines HS-27A (ATCC 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-5 supports proliferation of hematopoietic progenitor cells when co-cultured in serum-deprived media with no exogenous factors.


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 are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:9 is recommended
Medium Renewal: Everh 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 Fred Hutchinson Cancer Res. Cntr.
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

Basic Documentation
Other Documentation
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