| 產品名稱 |
HFF-1 |
| 商品貨號 |
B167200 |
| Organism |
Homo sapiens, human |
| Tissue |
Skin; foreskin |
| Cell Type |
Fibroblast |
| 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 |
Normal |
| Age |
Newborn |
| Gender |
male |
| Applications |
Can be used to produce feeder cells |
| Storage Conditions |
Liquid nitrogen vapor phase |
| Derivation |
The cell line was established by ATCC in 2003 from normal human foreskin pooled from two individuals. |
| Clinical Data |
male |
| Comments |
The growth of these cells should be arrested before being used as a feeder layer. ATCC has successfully irradiated (SCRC-1041.1) and treated the cells with Mitomycin C (SCRC-1041.2) for use as a feeder layer. If the HFFs are being used as a feeder layer for ES cells, it is not recommended to use them past passage no. 50 (P50). It is recommended that the feeder cells be plated 24 hours before use at 5X104 cells/cm2 in order to obtain a supportive monolayer for stem cell growth. |
| 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 15%
This medium is formulated for use with a 5% CO2 in air atmosphere. (Standard DMEM formulations contain 3.7 g/L sodium bicarbonate and a 10% CO2 in air atmosphere is then recommended).
|
| Subculturing |
Procedure:
To insure the highest level of viability, be sure to warm media and Trypsin/ EDTA to 37ºC before using it on the cells.
Cells should be split when they reach confluency. A split ratio of 1:5 to 1:7 is recommended. Volumes used in this protocol are for 225 cm2 (T225); proportionally reduce or increase amount of dissociation medium for culture flasks of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 1X PBS (SCRR-2201) solution to remove all traces of serum, which contain trypsin inhibitor.
- Add 5 mL of Trypsin-EDTA (0.25% (w/v) Trypsin-0.53 mM EDTA solution, ATCC# 30-2101) solution to flask and incubate for 1 minute, gently tapping the flask observe cells under an inverted microscope until cells detach (usually within 1 to 2 minutes).
- Add 6.0 to 8.0 mL of complete growth medium and rinse surface of the flask to detach all cells. Gently pipetting up and down will break cell clumps.
- Transfer all cells into a centrifuge bottle or tube and centrifuge at 270 xg for 5 minutes.
- Remove and discard the supernatant
- Add 10 mL complete growth medium to cell pellet and with 10 mL pipette resuspend the cells gently (create a single-cell suspension).
- Add more complete growth medium to cell suspension as needed to plate cells at approximately 5x106/T225 flask.
- Place flasks in incubator @ 37°C with a 5% CO2 in air atmosphere.
| Cryopreservation |
Growth Area (cm2)
|
| Culture Conditions |
1xPBS (mL)
|
| Name of Depositor |
Trypsin/EDTA (mL)
|
| Year of Origin |
Equal vol. Complete Growth Medium (mL)
|
| References |
Growth Medium (mL)
|
| 梅經理 |
17280875617 |
1438578920 |
| 胡經理 |
13345964880 |
2438244627 |
| 周經理 |
17757487661 |
1296385441 |
| 于經理 |
18067160830 |
2088210172 |
| 沈經理 |
19548299266 |
2662369050 |
| 李經理 |
13626845108 |
972239479 |
|
