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MEF (CF-1) IRR

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  • 2025年11月27日
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    • 文献和实验
    • 技术资料
    • 物种来源：

      小鼠

    • 是否是肿瘤细胞：

      0

    • 细胞类型：

      成纤维细胞

    • 细胞形态：

      成纤维样

    • ATCC Number：

      SCRC-1040.1™

    • 年限：

      14 days gestation embryo

    • 器官来源：

      胚胎

    • 生长状态：

      贴壁生长

    • 库存：

      大量

    • 运输方式：

      冻存运输

    Designations: MEF (CF-1) IRR
    Depositors:  ATCC
    Biosafety Level: 1
    Shipped: frozen
    Medium & Serum: See Propagation
    Growth Properties: adherent
    Organism: Mus musculus
    Morphology: fibroblast
    Source: Organ: embryo
    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.
    Isolation: Isolation date: 2004
    Applications: Irradiated cells for use as feeder layer
    Age: 14 days gestation embryo
    Gender: male and female mixed
    Comments: 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 10,000 rads and will not replicate. The cells will begin to deteriorate in 7 to 10 days 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 4 to 5 X 10(6) cells per T75 flask in order to obtain a 100% confluent monolayer for stem cells growth.
    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. Flasks do not need to be coated before plating MEFs.
    1. 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.
    2. 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.
    3. Transfer the vial s contents plus 5 ml of complete medium (see below for recipe) 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.
    4. Gently mix and pellet the cells by centrifugation @ 270 xg for 5 minutes.
    5. Discard the supernatant and resuspend the cells with 10ml fresh growth medium (warm) and transfer to one T75 flask.
    6. Add 5 ml more fresh growth medium (warm) to flask.
    7. Incubate 37�C in a 5% CO2 in air atmosphere.
    8. Fluid change twice a week or when pH decreases. 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).
    Cells should be plated 24 hours before use as a feeder layer for ES cells and kept for no more than 7 days.
    Medium Renewal: Twice a week or when 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-1040
    References: 89421: Nagy A, et al. Manipulating The Mouse Embryo: A Laboratory Manual. Third Edition: Cold Spring Harbor Press; 2003.

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    图标文献和实验
    相关实验

    • MEF小鼠胚胎成纤维细胞富集/消化/传代知识总结

      ，这发生在准备胚胎的第二天。这可能或早或晚发生，所以请注意观察你的培养瓶。 注释： 我们已用CF-1品系的鼠制备了成纤维细胞。 培养基成分： 88% DMEM 10% FBS 1% NEAA 1% 双抗 对于新建立的细胞系，要对样本进行支原体检测。 小鼠胚胎成纤维细胞的消化/传代 1、移去MEF培养基 2、用5ml不含钙镁离子的PBS洗涤细胞(以去除胰蛋白酶的抑制

    • MEF小鼠胚胎成纤维细胞富集/消化/传代知识总结

      时期。一般说来，这发生在准备胚胎的第二天。这可能或早或晚发生，所以请注意观察你的培养瓶。 注释： 我们已用CF-1品系的鼠制备了成纤维细胞。 培养基成分： 88% DMEM 10% FBS 1% NEAA 1% 双抗 对于新建立的细胞系，要对样本进行支原体检测。 小鼠胚胎成纤维细胞的消化/传代 1、移去MEF培养基 2、用5ml不含钙镁离子的PBS洗涤细胞(以去除胰蛋白酶的抑制

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