IMR-90細(xì)胞(IMR-90人胚肺成纖維細(xì)胞)ATCC® CCL-186™ 詳細(xì)資料
基本信息?
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ATCC編號(hào):CCL-186™
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? designation:IMR-90
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? depositors:WW Nichols(W·W·尼科爾斯)
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生物安全等級(jí):1級(jí)(BSL-1)
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運(yùn)輸方式:冷凍(干冰)
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生長(zhǎng)特性:貼壁生長(zhǎng)(adherent)
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種屬:人(Homo sapiens)
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形態(tài):成纖維細(xì)胞樣(fibroblast)
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組織來(lái)源:肺(lung)
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疾病狀態(tài):正常(normal)
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細(xì)胞類型:成纖維細(xì)胞(fibroblast)
許可與法規(guī)?
除上述內(nèi)容提及的材料轉(zhuǎn)讓協(xié)議(MTA)外,轉(zhuǎn)移該ATCC材料可能需要其他ATCC和/或監(jiān)管許可。購(gòu)買ATCC材料的用戶最終負(fù)責(zé)獲取此類許可。如需了解運(yùn)往您所在地區(qū)的具體要求,請(qǐng)點(diǎn)擊此處查詢。
應(yīng)用?
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轉(zhuǎn)染宿主(適用于Lonza公司的Nucleofection技術(shù))。
病毒易感性?
該細(xì)胞對(duì)以下病毒敏感:
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脊髓灰質(zhì)炎病毒1型(Human poliovirus 1)
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脊髓灰質(zhì)炎病毒2型(Human poliovirus 2)
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水痘-帶狀皰疹病毒(Varicella-Zoster)
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單純皰疹病毒1型(Herpes simplex virus 1)
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單純皰疹病毒2型(Herpes simplex virus 2)
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脊髓灰質(zhì)炎病毒3型(Human poliovirus 3)
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痘苗病毒(Vaccinia virus)
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巨細(xì)胞病毒(Human herpesvirus 5,CMV)
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水泡性口炎病毒(Vesicular stomatitis virus)
分子特征?
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反轉(zhuǎn)錄酶活性:陰性(Reverse Transcript: negative)
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STR DNA譜型(短串聯(lián)重復(fù)序列):
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Amelogenin(性別位點(diǎn)):X
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CSF1PO:11,13
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D13S317:11,13
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D16S539:10,13
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D5S818:12,13
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D7S820:9,12
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THO1:8,9.3
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TPOX:8,9
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vWA:16,19
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細(xì)胞遺傳學(xué)分析:正常女性二倍體(46,XX),染色體穩(wěn)定(normal human female; diploid; stable)
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同工酶:葡萄糖-6-磷酸脫氫酶(G6PD)類型為B(Isoenzymes: G6PD, B)
來(lái)源與背景?
IMR-90細(xì)胞系由W·W·Nichols及其同事從16周齡女性胎兒的正常肺組織中分離得到,是一株人二倍體成纖維細(xì)胞。
該細(xì)胞的分裂潛能、病毒易感性及其他特性已被深入研究,可作為WI-38(另一常用人肺成纖維細(xì)胞株)及其他標(biāo)準(zhǔn)人肺細(xì)胞株的替代選擇。
據(jù)報(bào)道,IMR-90細(xì)胞在出現(xiàn)衰老前的群體倍增次數(shù)(PDL)約為58次。
培養(yǎng)條件?
1. 完全生長(zhǎng)培養(yǎng)基?
基礎(chǔ)培養(yǎng)基為ATCC配制的Eagle必需培養(yǎng)基(Eagle's Minimum Essential Medium),貨號(hào)30-2003。
配制完全培養(yǎng)基時(shí),需向基礎(chǔ)培養(yǎng)基中添加10%胎牛血清(FBS)。
2. 培養(yǎng)環(huán)境?
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氣體氛圍:空氣(95%)+ 二氧化碳(CO?,5%)
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溫度:37.0°C
3. 傳代(Subculturing)?
操作步驟:
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移除并丟棄舊培養(yǎng)液。
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用0.25%(w/v)胰蛋白酶-0.53 mM EDTA溶液短暫沖洗細(xì)胞層,以去除含胰酶抑制劑的血清殘留。
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向培養(yǎng)瓶中加入2.0-3.0 ml胰蛋白酶-EDTA溶液,在倒置顯微鏡下觀察,直至細(xì)胞層分散(通常需5-15分鐘)。
注意:避免搖晃或敲擊培養(yǎng)瓶,防止細(xì)胞結(jié)團(tuán)。難以分散的細(xì)胞可置于37°C培養(yǎng)箱中加速解離。
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加入6.0-8.0 ml完全生長(zhǎng)培養(yǎng)基,輕輕吹打細(xì)胞以終止胰酶消化。
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將細(xì)胞懸液分裝至新的培養(yǎng)容器(如培養(yǎng)瓶或培養(yǎng)皿)。
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置于37°C、5% CO?培養(yǎng)箱中培養(yǎng)。
傳代比例:推薦1:2至1:8(即1份細(xì)胞懸液分至2-8個(gè)新容器)。
培養(yǎng)基更換頻率:每3-4天更換一次新鮮完全培養(yǎng)基。
4. 凍存(Preservation)?
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凍存液:完全生長(zhǎng)培養(yǎng)基(95%)+ 二甲基亞砜(DMSO,5%)。
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儲(chǔ)存溫度:液氮蒸氣相(liquid nitrogen vapor phase)。
相關(guān)產(chǎn)品?
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推薦基礎(chǔ)培養(yǎng)基(不含額外補(bǔ)充劑或血清):ATCC 30-2003(Eagle's Minimum Essential Medium)。
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推薦血清:ATCC 30-2020(胎牛血清)。
參考文獻(xiàn)?
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Nichols WW, et al. Characterization of a new human diploid cell strain, IMR-90. Science196: 60-63, 1977.(PubMed: 841339)
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Dolganov GM, et al. Human Rad50 is physically associated with human Mre11: identification of a conserved multiprotein complex implicated in recombinational DNA repair. Mol Cell Biol16: 4832-4841, 1996.(PubMed: 8756642)
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Ostlund RE Jr., et al. A stereospecific myo-inositol/D-chiro-inositol transporter in HepG2 liver cells. J Biol Chem271: 10073-10078, 1996.(PubMed: 8626564)
其他信息?
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細(xì)胞庫(kù)信息:中國(guó)科學(xué)院典型培養(yǎng)物保藏委員會(huì)細(xì)胞庫(kù)(GNHu32)提供該細(xì)胞,經(jīng)支原體、細(xì)菌、真菌檢測(cè)均為陰性,且通過(guò)STR鑒定無(wú)誤。
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供應(yīng)限制:僅供研究使用(For research use only)。
IMR-90細(xì)胞, 人胚肺成纖維細(xì)胞
ATCC® Number: CCL-186™
Designations: IMR-90
Depositors: WW Nichols
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: fibroblast
Source: Organ: lung
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 ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications: transfection host (Nucleofection technology from Lonza)
Virus Susceptibility: Human poliovirus 1
Human poliovirus 2
Varicella-Zoster
Herpes simplex virus 1
Herpes simplex virus 2
Human poliovirus 3
Vaccinia virus
Human herpesvirus 5
Vesicular stomatitis virus
Reverse Transcript: negative
DNA Profile (STR): Amelogenin: X
CSF1PO: 11,13
D13S317: 11,13
D16S539: 10,13
D5S818: 12,13
D7S820: 9,12
THO1: 8,9.3
TPOX: 8,9
vWA: 16,19
Cytogenetic Analysis: normal human female; diploid; stable
Isoenzymes: G6PD, B
Age: 16 weeks gestation
Gender: female
Ethnicity: Caucasian
Comments: The human diploid fibroblast strain IMR-90 was derived by W.W. Nichols and associates from the lungs of a 16-week female fetus. [22381]
The division potential, viral susceptibilities and other properties have been thoroughly studied such that the line may be considered as an alternate for WI-38 and other standard human lung cell strains.
The cells have been reported to be capable of attaining 58 population doublings before the onset of senescence. [22381]
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells 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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subc*tion Ratio: A subc*tion ratio of 1:2 to 1:8 is recommended
Medium Renewal: Every 3 to 4 days
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003
recommended serum:ATCC 30-2020
References: 22381: Nichols WW, et al. Characterization of a new human diploid cell strain, IMR-90. Science 196: 60-63, 1977. PubMed: 841339
32932: Dolganov GM, et al. Human Rad50 is physically associated with human Mre11: identification of a conserved multiprotein complex implicated in recombinational DNA repair. Mol. Cell. Biol. 16: 4832-4841, 1996. PubMed: 8756642
33041: Ostlund RE Jr., et al. A sterospecific myo-inositol/D-chiro-inositol transporter in HepG2 liver cells. J. Biol. Chem. 271: 10073-10078, 1996. PubMed: 8626564
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