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BE(2)-M17细胞，人神经母细胞瘤细胞

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  • CRL-2237
  • 2025年11月18日
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    BE(2)-M17人神经母细胞瘤细胞 ATCC 细胞|细胞系|细胞株|肿瘤细胞|细胞|贴壁细胞|悬浮细胞|。细胞库管理规范，提供的细胞株背景清楚，提供参考文献和培养条件!...

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    BE(2)-M17人神经母细胞瘤细胞
    ATCC® Number: CRL-2237™ Price: $329.00
    Designations: BE(2)-C
    Depositors: JL Biedler
    Biosafety Level: 1
    Shipped: frozen
    Medium & Serum: See Propagation
    Growth Properties: adherent
    Organism: Homo sapiens (human)
    Morphology: neuroblast
    Source: Organ: brain
    Disease: neuroblastoma
    Derived from metastatic site: bone marrow
    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: BE(2)-C was deposited at the ATCC by June L. Biedler, Memorial Sloan-Kettering Cancer Center. BE(2)C is distributed for academic research purposes only. Memorial Sloan-Kettering releases the line subject to the following: 1.)BE(2)-C or its products must not be distributed to third parties. Commercial interests are the exclusive property of Memorial Sloan-Kettering Cancer Center. 2.) Any proposed commercial use of BE(2)-C including any use by a for-profit entity must first be negotiated with Director, Office of Industrial Affairs, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021; phone (212) 639-6181; FAX (212) 717-3439.
    Isolation: Isolation date: November, 1972
    DNA Profile (STR): Amelogenin: X
    CSF1PO: 10
    D13S317: 11
    D16S539: 9,11
    D5S818: 12
    D7S820: 9,10
    THO1: 6
    TPOX: 11
    vWA: 18
    Age: 2 years
    Gender: male
    Comments: BE(2)-C is a clone of the SK-N-BE(2) neuroblastoma cell line (see ATCCCRL-2271) that was established in November of 1972 from a bone marrow biopsy taken from child with disseminated neuroblastoma after repeated courses of chemotherapy and radiotherapy.
    BE(2)-C cells have a reported saturation density of greater than 5 X 10 exp5 cells/sq cm.
    The cells grow as clusters of flattened neuroblastic cells with occasional fine cell processes (neurites).
    Unlike the parent line, they generally do not detach and float.
    Propagation: ATCC complete growth medium: The base medium for this cell line is a 1:1 mixture of ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003, and F12 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
    Temperature: 37.0°C
    Subculturing: Protocol:
    1. Remove and discard culture medium.
    2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
    3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask anells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 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.
    4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
    5. Add appropriate aliquots of the cell suspension to new culture vessels.
    6. Incubate cultures at 37°C.

    Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
    Medium Renewal: Every 2 to 3 days
    Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%
    Storage temperature: liquid nitrogen vapor phase
    Doubling Time: 18 hrs
    Related Products: recommended serum:ATCC 30-2020
    parental cell line:ATCC CRL-2271

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    二、细胞收到后处理

    细胞在培养瓶中培养至良好状态后灌满完全培养液并封好瓶口是细胞运输的最好办法。收到细胞回到自己的实验室后，先打开外包装，用75%酒精喷洒整个瓶消毒后放到超净台内，严格无菌操作，培养箱静置2-4小时。镜下观察：未超过80%汇合度时，可将瓶装的完全培养液收集至离心管中，加入6ml完全培养基，放入37℃、5%CO2孵箱培养；超过80%汇合度时，根据情况传代或者冻存，具体操作见细胞培养步骤。（注意发货的是密封培养瓶的话，处理完后放入培养箱培养记得培养瓶盖子拧松，传代后建议一瓶用原瓶里面的完全培养基，另外一瓶用自己配的完全培养基）

    三、细胞培养步骤
     
    1. 复苏细胞：将含有1mL细胞悬液的冻存管在37℃水浴中迅速摇晃解冻，加入5mL培养基混合均匀。在1000RPM条件下离心5分钟，弃去上清液，补加4-6mL完全培养基后吹匀。然后将所有细胞悬液加入培养瓶中培养过夜（或将细胞悬液加入6cm皿中），培养过夜。第二天换液并检查细胞密度。
    2. 细胞传代：如果细胞密度达80%-90%，即可进行传代培养。

    1、对于贴壁细胞，传代可参考以下方法：
    1. 弃去培养上清，用不含钙、镁离子的PBS润洗细胞1-2次。
    2. 加1-2ml消化液（0.25%Trypsin-0.53mM EDTA）于培养瓶中，置于37℃培养箱中消化1-2min，然后在显微镜下观察细胞消化情况，若细胞大部分变圆并脱落，迅速拿回操作台，轻敲几下培养瓶后加5ml以上含10%血清的完全培养基终止消化。
    3.轻轻吹打细胞，完全脱落后吸出，在1000RPM条件下离心8-10分钟，弃去上清液，补加1-2mL培养液后吹匀。
     
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    BE(2)-M17细胞，人神经母细胞瘤细胞
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