hct-116、hela、MEFs细胞参数介绍.doc

HELAHeLa (ATCC® CCL-2™)

ATCC：Hela细胞，源于黑人31岁女性，子宫颈腺癌，是一种附着型上皮细胞，要求存于液氮中。These cells are a suitable transfection host.This cell line can be used to screen for Escherichia coli strains with invasive potential.
Biosafety Level：（2 [Cells contain human papilloma virus]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.）
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%.
Subculturing（接种）：Volumes used in this protocol are for a 75 cm2flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. CorningT-75 flasks (catalog #430641) are recommended for subculturing this product.
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 30 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 37C 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 37C.
Subcultivation Ratio:A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal:2 to 3 times per week
Cryopreservation
Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C

论坛：贴壁生长，铺路石状长得快，2-3天传代一次，传代不及时会造成老化的细胞堆积，看起来很脏。10%+1640或者10%+高糖DMEM都有培养。

MEFs_283TAg (ATCC® CRL-2822™)
Organism
Mus musculus, mouse
Tissue
embryo
Cell Type
fibroblast immortalized with SV40 large T antigenSV40 large T antigen transfected
Product Format
frozen
Morphology
fibroblast
Culture Properties
adherent
Biosafety Level
2 cells containing SV40 viral DNA sequences

Biosafety classification is based on聽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.
Age
14.5 day gestation embryo
Applications
DNA repair studies
Storage Conditions
liquid nitrogen vapor phase
Derivation
283TAg (Polb/Mpg double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/ N-methylpurine-DNA glycosylase (Mpg, Aag) double null embryos at day 14.5 of gestation. The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII).
Comments
283TAg (Polb/Mpg double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/ N-methylpurine-DNA glycosylase (Mpg, Aag) double null embryos at day 14.5 of gestation. The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII).
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 10%. 
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 that contains trypsin inhibitor.
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 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.
An inoculum of 4 X 10(3) to 4 X 10(4) viable cells/cm2 is recommended.
6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 6 X 10(3) and 1 X 10(5) cells/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Two to three times weekly
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Population Doubling Time
29 hours

HCT 116 (ATCC® CCL-247™)
Organism
Homo sapiens, human
Tissue
colon
Product Format
frozen
Morphology
epithelial
Culture Properties
adherent
Biosafety Level
1

Biosafety classification is based on聽U.S. Public Health Service Guidelines,聽it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease
colorectal carcinoma(结肠直肠癌)
Age
adult
Gender
male
Applications
This cell line is a suitable transfection host.
This line has a mutation in codon 13 of the ras proto-oncogene, and can be used as a positive control for PCR assays of mutation in this codon.
Storage Conditions
liquid nitrogen vapor phase

Karyotype
The stemline chromosome number is near diploid with the modal number at 45 (62%) and polyploids occurring at 6.8%. The markers 10q+ and t(?8p;18q) are present in all metaphases and t(9q;?16p-), in 80% of the cells karyotyped. N16 is monosomic in the presence of, but disomic in the absence of t(9q;?16p-). N10 and N18 are monosomic and other chromosomes from those mentioned above are disomic. Q-band observations revealed the presence of the Y chromosome, but not in all cells (50% of cells lacked the Y in G-band karyotypes).
Images

Clinical Data
male
Genes Expressed
carcinoembryonic antigen (CEA) 1 ng per 106聽cells per 10 days.
Cellular Products
carcinoembryonic antigen (CEA) 1 ng per 10 exp6 cells per 10 days; keratin
Tumorigenic
Yes
Effects
Yes, in nude mice
Ref
Comments
The cells are positive for keratin by immunoperoxidase staining.
HCT 116 cells are positive for transforming growth factor beta 1 (TGF beta 1) and beta 2 (TGF beta 2) expression.
Complete Growth Medium
The base medium for this cell line is ATCC-formulated McCoy's 5a Medium Modified, Catalog No. 30-2007. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.聽
Subculturing
Volumes are given for a 75 cm2聽flask. Corning庐聽T-75 flasks (catalog #430641) are recommended for subculturing this product. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
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 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掳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掳C.
Subcultivation Ratio:聽A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal:聽Every 2 to 3 days
Cryopreservation
Freeze medium:聽Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature:聽liquid nitrogen vapor phase
Culture Conditions
Atmosphere:聽air, 95%; carbon dioxide (CO2), 5%
Temperature:聽37掳C
Growth Conditions:聽Growth and plating efficiency are enhanced by using a feeder layer of murine fibroblasts.

人结肠癌ＨＣＴ－ １１６细胞是Ｂｒａｔｔａｉｎ等人在１９７９年从一例男性结肠癌患者的癌组织中培养建立起来的低分化型 腺癌细胞株［１］ ，广泛应用于恶性肿瘤细胞的生物学特性、抗 肿瘤药物作用机理和抗癌药物的筛选等领域的研究 。近年的研究表明，大多数ＨＣＴ－ １１６细胞具有肿瘤干细胞的特性，可作为肿瘤干细胞研究的理想对象［ ６］ 。在体外培养条件下，ＨＣＴ－ １１６细胞一般呈梭形或多角形贴壁生长，传代培养时通常采用胰蛋白酶消化法进行。