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?

• ATCC編號：CCL-186™
• ? designation：IMR-90
• ? depositors：WW Nichols（W·W·尼科爾斯）
• 生物安全等級：1級（BSL-1）
• 運(yùn)輸方式：冷凍（干冰）
• 生長特性：貼壁生長（adherent）
• 種屬：人（Homo sapiens）
• 形態(tài)：成纖維細(xì)胞樣（fibroblast）
• 組織來源：肺（lung）
• 疾病狀態(tài)：正常（normal）
• 細(xì)胞類型：成纖維細(xì)胞（fibroblast）

許可與法規(guī)?

應(yīng)用?

• 轉(zhuǎn)染宿主（適用于Lonza公司的Nucleofection技術(shù)）。

病毒易感性?

• 脊髓灰質(zhì)炎病毒1型（Human poliovirus 1）
• 脊髓灰質(zhì)炎病毒2型（Human poliovirus 2）
• 水痘-帶狀皰疹病毒（Varicella-Zoster）
• 單純皰疹病毒1型（Herpes simplex virus 1）
• 單純皰疹病毒2型（Herpes simplex virus 2）
• 脊髓灰質(zhì)炎病毒3型（Human poliovirus 3）
• 痘苗病毒（Vaccinia virus）
• 巨細(xì)胞病毒（Human herpesvirus 5，CMV）
• 水泡性口炎病毒（Vesicular stomatitis virus）

分子特征?

• 反轉(zhuǎn)錄酶活性：陰性（Reverse Transcript: negative）
• STR DNA譜型（短串聯(lián)重復(fù)序列）：
  • Amelogenin（性別位點(diǎn)）：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
• 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）
• 同工酶：葡萄糖-6-磷酸脫氫酶（G6PD）類型為B（Isoenzymes: G6PD, B）

來源與背景?

培養(yǎng)條件?

1. 完全生長培養(yǎng)基?

2. 培養(yǎng)環(huán)境?

• 氣體氛圍：空氣（95%）+ 二氧化碳（CO?，5%）
• 溫度：37.0°C

3. 傳代（Subculturing）?

1. 移除并丟棄舊培養(yǎng)液。
2. 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。
3. 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。
4. 加入6.0-8.0 ml完全生長培養(yǎng)基，輕輕吹打細(xì)胞以終止胰酶消化。
5. 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）。
6. 置于37°C、5% CO?培養(yǎng)箱中培養(yǎng)。

4. 凍存（Preservation）?

• 凍存液：完全生長培養(yǎng)基（95%）+ 二甲基亞砜（DMSO，5%）。
• 儲存溫度：液氮蒸氣相（liquid nitrogen vapor phase）。

相關(guān)產(chǎn)品?

• 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）。
• 推薦血清：ATCC 30-2020（胎牛血清）。

參考文獻(xiàn)?

1. Nichols WW, et al. Characterization of a new human diploid cell strain, IMR-90. Science196: 60-63, 1977.（PubMed: 841339）
2. 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）
3. 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?

• 細(xì)胞庫信息：中國科學(xué)院典型培養(yǎng)物保藏委員會細(xì)胞庫（GNHu32）提供該細(xì)胞，經(jīng)支原體、細(xì)菌、真菌檢測均為陰性，且通過STR鑒定無誤。
• 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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