HULEC-5a NTCC®人肺微血管内皮细胞株及完全培养基-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心 CRL-3244

HULEC-5a cell line细胞株及完全培养基-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

HULEC-5a Cell Line

Cat No.: NTCC-CRL3244

Organism Homosapiens, human

Tissue lung

Cell Typemicrovascularendothelium

Product Formatfrozen

Morphology endothelial-like

Culture Properties adherent

Biosafety Level 2[Cells contain SV-40 viral DNA sequences]

Agenewborn

Gendermale

ApplicationsInvestigatingthe role of endothelium in infection by virus and bacteria, cancer research,tumor metastastis, endothelium function, cell trafficking, surface moleculeinteractions

Derivation

Microvascular endothelial cells isolated from humanlungs were transfected with pSVT vector,a pBR-322 based plasmid containing the coding region for Simian virus40A gene product, large T antigen.

Receptor ExpressionneonatalFcRn receptor (FcRn) (Goebl N, et al. Neonatal Fc Receptor MediatesInternalization of Fc in Transfected Human Endothelial Cells. Mol. Biol. Cell19: 5490-5505, 2008. PubMed: 18843053)

Comments CRL-3244,HULEC-5a, has been shown to retain many of the characteristics of endothelialcells. These immortalized cells, because they are a continously renewablesource of human endothelial microvascular cells, can be used as a replacementfor primary human lung endothelial cells for many research studies.

Complete Growth MediumThe base medium for this cell line is MCDB131 (withoutL-Glutamine). To make the complete growth medium, add the following componentsto the base medium:

10ng/mL Epidermal Growth Factor (EGF)

1 µg/mL Hydrocortisone

10 mM Glutamine

fetal bovine serum (FBS) to a final concentration of10%

SubculturingVolumesused in this protocol are for 75 cm2 flasks; proportionally reduce or increaseamount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium. Briefly rinse thecell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (NTCC30-2200) or 0.25% Trypsin –0.53mM EDTA (NTCC 30-2101) solution to remove all traces of serum whichcontains trypsin inhibitor.

Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flaskand 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 byhitting or shaking the flask while waiting for the cells to detach. Cells thatare difficult to detach may be placed at 37°C to facilitate dispersal.

Add 6.0 to 8.0 mL of complete growth medium andaspirate cells by gently pipetting.

Add appropriate aliquots of the cell suspension tonew culture vessels. Incubate cultures at 37°C.

Subcultivation Ratio: 1:5 to 1:10 is recommended.

Medium Renewal: Do not feed cells with growth mediumbetween subcultures or cells become very tightly attached and will be difficultto disperse with trypsin-EDTA.

Cryopreservation

Freeze Medium: Complete growth medium, 92.5%; DMSO,7.5%

Storage Temperature: liquid nitrogen vapor phase

Culture ConditionsTemperature:37°C

STR ProfileAmelogenin:X

CSF1PO: 11, 12

D13S317: 13, 14

D16S539:9,10

D5S818: 11, 13

D7S820: 8, 10

TH01: 6, 7

TPOX: 8, 11

vWA: 16, 17

Year of OriginNovember1990

References

Lawrence EC et al. Establishment of immortalizedhuman lung microvascular endothelial cell lines. Chest 108(3) Supplement:126S,1995.

【Organism物种来源】人源Homo sapiens, human  
【Tissue组织来源】
【Cell Type细胞类型】  
【Product Format产品状态】 frozen/live culture
【Morphology细胞形态】  
【Culture Properties细胞特性】
【Biosafety Level生物安全级别】 1/2
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.

【Disease细胞源疾病】
【Age年龄】
【Gender性别】 Male/Female
【Storage Conditions保存条件】 liquid nitrogen vapor phase
【Karyotype染色体组型】
【Images细胞图片】 Cell Micrograph
【Derivation衍生细胞】 ­
【Clinical Data临床数据】
【Comments其他描述】
【Complete Growth Medium完全培养基】 The base medium for this cell line is DMEM/RPMI-1640 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
【Subculturing传代方法】
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
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.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
【Cryopreservation冻存条件】
【Freeze Medium冻存培养基】Complete growth medium supplemented with 5% (v/v) DMSO
【Storage Temperature保存温度】 Liquid nitrogen vapor phase
【Culture Conditions培养条件】
【Atmosphere培养环境】 Air, 95%; carbon dioxide (CO2), 5%
【Temperature培养温度】 37°C
【STR Profile图谱】
【Population Doubling Time倍增殖时间】
【References参考文献】

【Supplier细胞供应厂家】BioVector NTCC Inc.
【Website网址】http://www.biovector.net