HFF-1 IRR細(xì)胞

ATCC Number： SCRC-1041.1?

運(yùn)輸方式： 凍存運(yùn)輸

相關(guān)**： 正常

器官來(lái)源： 皮膚

年限： newborn

數(shù)量： 大量

細(xì)胞類型： 成纖維細(xì)胞

細(xì)胞形態(tài)： 成纖維樣

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 人

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

Designations: HFF-1 IRR

Depositors: ATCC

Biosafety Level: 1

HFF-1 IRR細(xì)胞Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: fibroblast

Source: Organ: skin; foreskin

Disease: normal

Cell Type: fibroblast

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Restrictions: Prior to purchase, for-profit commercial institutions must obtain a license agreement. For instructions on how to proceed, please contact ATCC 's Office of Licensing and Business Development at licensing@ATCC .org or 703 365 2773.

Isolation: Isolation date: 2003

Applications: Irradiated cells for use as feeder layer

Age: newborn

Gender: male

Comments: HFF-1 IRR細(xì)胞These cells are provided to be used as feeder cells to support the growth of stem cells in the undifferentiated state. They have been irradiated with 6,000 rads and will not replicate. The cells will begin to deteriorate in 2 to 3 weeks after plating. Once the feeder cells have attached, the culture medium can be changed to accommodate the cells to be supported. It is recommended that the feeder cells be plated 24 hours before use at 5 to 6 X 10(6)/T75 in order to obtain a 100% confluent monolayer for stem cells growth. It is not recommended to use them past passage no. 50 (P50).

Propagation: ATCC 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%

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Subculturing: Protocol: Establishing and maintaining your culture:

To insure the highest level of viability, be sure to warm media to 37?C before using it on the cells. Volumes used in this protocol are for T75 flasks. Proportionally adjust the volumes for culture vessels of other sizes.

Thaw the vial by gentle agitation in a 37?C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.

Remove the vial from the water bath as soon as the contents are half way thawed (approximately 90 seconds), and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

Transfer the vial s contents plus 5 ml of complete medium to a 15 ml centrifuge tube. Use an additional 1 ml of medium to rinse the vial and transfer the liquid to the 15 ml tube. Add 4 ml of complete medium to bring the total volume to 10 ml.

Gently mix and pellet the cells by centrifugation @ 270 xg for 5 minutes.

Discard the supernatant and resuspend the cells with 10ml fresh growth medium (warm) and transfer to one coated T75 flask.

Incubate 37?C in a 5% CO2 in air atmosphere.

Fluid change twice a week or when pH decreases. HFF-1 IRR細(xì)胞It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

Medium Renewal: Twice a week or as pH decreases

Preservation: Storage temperature: liquid nitrogen vapor phase

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

Recommended serum: ATCC 30-2020

Source culture: ATCC SCRC-1041

References: 89580: Amit M, et al. Human feeder layers for human embryonic stem cells. Biol. Reprod. 68: 2150-2156, 2003. PubMed: 12606388

89581: Hovatta O, et al. A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells. Hum. Reprod. 18: 1404-1408, 2003. PubMed: 12832363

89586: Andrews P, et al. HFF-1 IRR細(xì)胞Human embryonic fibroblast feeder cells. International Patent Application WO 03/078611 A1