HFF-1 IRR细胞
ATCC Number： SCRC-1041.1™
运输方式： 冻存运输
相关**： 正常
器官来源： 皮肤
年限： newborn
数量： 大量
细胞类型： 成纤维细胞
细胞形态： 成纤维样
是否是肿瘤细胞： 0
物种来源： 人
生长状态： 贴壁生长
Designations: HFF-1 IRR
Depositors: ATCC
Biosafety Level: 1
HFF-1 IRR细胞Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens
Morphology: fibroblast
Source: Organ: skin; foreskin
Disease: normal
Cell Type: fibroblast
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Isolation: Isolation date: 2003
Applications: Irradiated cells for use as feeder layer
Age: newborn
Gender: male
Comments: HFF-1 IRR细胞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细胞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细胞Human embryonic fibroblast feeder cells. International Patent Application WO 03/078611 A1
