| 產(chǎn)品名稱 |
HS-27A |
| 商品貨號 |
B164787 |
| 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]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| 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, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) 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.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- 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 |
| Deposited As |
human |
| 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
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