| 產品名稱 |
L8 |
| 商品貨號 |
B164950 |
| Organism |
Rattus norvegicus, rat |
| Tissue |
skeletal muscle |
| Cell Type |
myoblast |
| Product Format |
frozen |
| Morphology |
fibroblast |
| Culture Properties |
adherent |
| Biosafety Level |
1
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. |
| Disease |
Carcinogen |
| Age |
newborn |
| Applications |
transfection host |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
This line was originally isolated by D. Yaffe in 1969 from primary rat skeletal muscle cultures.
Unlike the L6 cell line (ATCC CRL-1458) no carcinogen was used to establish the L8 line. |
| Genes Expressed |
creatine phosphokinase (CPK); myosin |
| Cellular Products |
creatine phosphokinase (CPK); myosin |
| Comments |
Upon becoming confluent, L8 will fuse to form cross striated multinucleated muscle fibers.
It is recommended that early passages be preserved in liquid nitrogen and that the line be recloned periodically and reselected for progeny that have the ability to fuse.
It is important that the cells be subcultured when the flask is about 60% confluent. The myoblastic population will be depleted if the cultures are allowed to become confluent since most of the cells will fuse into nondividing syncytia. To avoid this, one must subculture before before the cultures become confluent and should reclone periodically and select myoblastic clones. |
| Complete Growth Medium |
A 4:1 mixture of Dulbecco's modified Eagle's medium and Medium 199, 89%; chicken embryo extract (US Biological cat# C3999), 1%; horse serum, 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.
- 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 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 at a density of 2000 cells/sq. cm.
- Incubate cultures at 37°C.
Subcultivation Ratio: An inoculation density of 4000 viable cells per cm2 of flask or dish surface area is recommended.
Medium Renewal: Twice per 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% |
| Name of Depositor |
B Paterson |
| Deposited As |
Rattus sp. |
| Passage History |
It is recommended that early passages be preserved in liquid nitrogen and that the line be recloned periodically and reselected for progeny that have the ability to fuse. |
| References |
Richler C, Yaffe D. The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev. Biol. 23: 1-22, 1970. PubMed: 5481965
Yaffe D, Saxel O. A myogenic cell line with altered serum requirements for differentiation. Differentiation 7: 159-166, 1977. PubMed: 558123
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