欧美精品AⅤ一区二区三区,亚洲乱码精品久久久久,99久久久国产精品免费蜜臀,精品人妻系列无码人妻免费视频,伊人久久精品无码麻豆一区,久久99人妻无码精品一区二区,国产精品免费久久久久影院仙踪林,久久99国产精品久久99果冻传媒

熱門搜索:A549    293T 金黃色葡萄球菌 大腸桿菌 AKK菌
購物車 1 種商品 - 共0元
當(dāng)前位置: 首頁 > ATCC代理 > 10.014 pRSV-T
最近瀏覽歷史
聯(lián)系我們
  • 0574-87157013
  • mingzhoubio@163.com
  • 浙江省寧波市鎮(zhèn)海區(qū)莊市街道興莊路9號
  • 創(chuàng)e慧谷42號樓B幢401室
10.014 pRSV-T
10.014 pRSV-T
規(guī)格:
貨期:
編號:B161387
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱 10.014 pRSV-T
商品貨號 B161387
Organism Homo sapiens, human
Tissue eye, cornea
Cell Type epithelial cells transfected with a plasmid containing the SV40 early region
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain SV-40 and Rous Sarcoma viral sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Storage Conditions liquid nitrogen vapor phase
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
Images
Derivation
A primary culture of normal corneal epithelium was immortalized by transfection with plasmid pRSV-T using lipofectamine overnight at 37°C. pRSV-T contains the SV40 early region genes and the Rous Sarcoma virus long terminal repeat.
Comments On a personal communication with the ATCC, the depositor commented that these cells have a finite life span.
Depositor Reference:
Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498.
Complete Growth Medium

The base medium for this cell line is Keratinocyte-Serum Free Medium (ThermoFisher cat # 17005-042) with 5 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract. To make the complete medium add the following components to the base medium: 0.005 mg/ml human recombinant insulin (ThermoFisher cat # 12585-014) and 500 ng/ml hydrocortisone (Sigma cat # H0135).

Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Note: The flasks used should be precoated with a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin. Subculture when 80% confluent.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.05% Trypsin-0.53 mM EDTA (ThermoFisher cat# 25300-054).
  3. 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 with 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  fresh medium and aspirate cells by gently pipetting. 
  5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium.  Add appropriate aliquots of cell suspension to new coated culture vessels. 
  7. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended when 80% confluent
Medium Renewal: Every 2 to 3 days.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.  


Cryopreservation
Freeze medium: complete growth medium supplemented with 7.5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X,Y
CSF1PO: 10
D13S317: 12,13
D16S539: 8,13
D5S818: 12,13
D7S820: 8,10
THO1: 6,8
TPOX: 8,10
vWA: 15
Name of Depositor CR Kahn, Gillette Medical Evaluation Laboratories
U.S. Patent Number
References

Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997

Kruszewski FH, et al. Evaluation of a human corneal epithelial cell line as an in vitro model for assessing ocular irritation. Fundam. Appl. Toxicol. 36: 130-140, 1997. PubMed: 9143482

梅經(jīng)理 17280875617 1438578920
胡經(jīng)理 13345964880 2438244627
周經(jīng)理 17757487661 1296385441
于經(jīng)理 18067160830 2088210172
沈經(jīng)理 19548299266 2662369050
李經(jīng)理 13626845108 972239479
新竹市| 庆安县| 罗田县| 顺平县| 金塔县| 江陵县| 喜德县| 油尖旺区| 霍城县| 高密市| 宁海县| 精河县| 南乐县| 徐水县| 高雄市| 砚山县| 安达市| 西林县| 德昌县| 沙湾县| 庄河市| 亳州市| 崇明县| 永修县| 清丰县| 西峡县| 清水河县| 合山市| 卓资县| 灵寿县| 万全县| 武冈市| 广丰县| 抚远县| 杭锦后旗| 西乡县| 疏附县| 汝南县| 岗巴县| 循化| 独山县|