| 產品名稱 |
PHM1-41 |
| 商品貨號 |
B192724 |
| Organism |
Homo sapiens, human |
| Tissue |
uterine myometrium smooth muscle |
| Cell Type |
fibroblast |
| Product Format |
frozen |
| Morphology |
fibroblast-like |
| Culture Properties |
adherent |
| Biosafety Level |
2 [cells contain Human Papilloma (HPV-16) viral DNA 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. |
| Gender |
female |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The PHM1-41 cell line was derived from pregnant human myometrium obtained at time of elective cesarean section, 39 weeks gestation, patient was not in labor, no complications in the pregnancy.
Myometrial cells were isolated by enzymatic digestion, infected at passage 2 with replication-defective adenovirus vector pLXSN16E6E7 expressing the E6/E7 proteins of human papilloma virus 16 and the neomycin resistance gene, and selected with the neomycin analog Geneticin. The designation -41 represent the subclones of the original immortalization mixture, selected by cloning rings for smooth muscle morphology but were not derived by limiting dilution cloning. |
| Clinical Data |
female patient, pregnant myometrium (39 weeks gestation) |
| Comments |
PHM1-41 cells retain many morphological and phenotypic responses in common with cultured primary myometrial cells. They are useful for long-term or coordinated studies,as their behavior is quite reproducible and the alternative, obtaining material from patients, is labor intensive and variable. These cells have been used to study crosstalk signaling between contractant and relaxant pathways, oxytocin receptor signaling pathways, and signal regulated control of calcium dynamics in the cytosol and endoplasmic reticulum compartments. Furthermore the can be used for studies on ion channel activity, expression, and gene regulation.
PHM1-41 does not grow in soft agar.
|
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, (ATCC® No. 30-2002). To make the complete growth medium, add the following components to the base medium:
- 0.1 mg/mL G-418
- additional 2 mM Glutamine
- heat-inactivated fetal bovine serum (FBS) to a final concentration of 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.
- Remove and discard culture medium.
- Briefly rinse the cell layer, twice, with 5 mL Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which containstrypsin inhibitor.
- Add 5 mL of 0.05% 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 5 mL of complete growth medium and aspirate cells by gently pipetting.
- Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
- Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new vessels. An inoculum of 1.0 X 14 to 2.0 X 104 viable cells/cm2 is recommended.
- Incubate cultures at 37°C.
Subcultivation ratio: A subcultivation ratio of 1:2 to 1:3 is recommended.
Medium renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze Medium: Complete growth medium, 95% supplemented with heat-inactivated fetal bovine serum + 5% DMSO |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| STR Profile |
TH01: 7
D5S818: 11, 13
D13S317: 14
D7S820: 8, 10
D16S539: 10, 12
CSF1PO: 11
Amelogenin: X
vWA: 16, 17
TPOX: 8, 11 |
| Sterility Tests |
Pass |
| Passage Number |
12 |
| Population Doubling Time |
43.2 hours |
| Name of Depositor |
B Sanborn |
| Year of Origin |
1992 |
| References |
Monga M, et al. Oxytocin-stimulated responses in a pregnant human immortalized myometrial cell line. Biol. Reprod. 55(2): 427-32, 1996. PubMed: 8828850
Yang M, et al. Multiple Trp isoforms implicated in capacitative calcium entry are expressed in human pregnant myometrium and myometrial cells. Biol Reprod 67(3):988-94, 2002. PubMed: 12193412
Zhong M., et al. Extracellular signal-regulated kinase 1/2 activation by myometrial oxytocin receptor involves Galpha(q)Gbetagamma and epidermal growth factor receptor tyrosine kinase activation. Endocrinology 144(7): 2947-56, 2003. PubMed: 12810550
Zhong M., et al. Multiple signals regulate phospholipase CBeta3 in human myometrial cells. Biol Reprod. 78(6): 1007-17, 2008. PubMed: 18322273
Murtazina DA, TRPC1, STIM1, and ORAI influence signal-regulated intracellular and endoplasmic reticulum calcium dynamics in human myometrial cells. Biol. Reprod. 85(2): 315-26, 2011. PubMed: 21565997
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