NE-GFP-4C cell line小鼠神經干細胞熒光表達細胞株/完全培養基-BioVector NTCC質粒載體菌種細胞蛋白抗體基因保藏中心 CRL-2926)

NE-GFP-4C

Cat# BioVector - CRL-2926

Product category

Animal cells

Organism

Mus musculus, mouse

Classification

Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod

Cell type

neural stem cell

Morphology

neuroepithelial

Tissue

Brain; Neuroectodermal

Applications

3D cell culture

Neuroscience

Growth properties

Adherent

Derivation

The neuroepithelial cell lines, NE-4C (CRL-2925) and NE-GFP-4C (CRL-2926) were established from the cerebral vesicles of 9-day-old mouse embryos lacking the functional p53 genes.

Age

embryo

Strain

C57Bl/Sv129 (p53-/-)

Antigen expression

Sox-2, Otx-2, En-1

Comments

Both NE-4C (CRL-2925) and NE-GFP-4C (CRL-2926) differentiate into neurons and astrocytes when exposed to retinoic acid. The GFP-transfected clone, NE-GFP-4C, when implanted into the forebrain of adult, new-born, and fetal mice or into the mid- and forebrain vesicles of early chick embryos is capable of developing morphologically differentiated neurons. Ref Ref

Handling information

Unpacking and storage instructions

Check all containers for leakage or breakage.

Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below --130°C, preferably in liquid nitrogen vapor, until ready for use.

Complete 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:

additional 2mM L-Glutamine

fetal bovine serum (FBS) to a final concentration of 10%

Temperature

37°C

Atmosphere

95% Air, 5% CO2

Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

Note: The culture flasks should be pre-coated with 15μg/mL poly-L-lysine (Sigma Cat. #P-9155 or equivalent) at least 2 hours in advance.

Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).

Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 10 minutes.

Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm2 or a 75 cm2 poly-L-lysine coated culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product.

Subculturing procedure

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

Note: The culture flasks should be pre-coated with15 μg/mL poly-L-lysine (Sigma Cat #P-9155) at least 2 hours in advance.

Remove and discard culture medium.

Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new poly-L-lysine coated culture vessels. An inoculum of 2 X 104 to 4 X 104 viable cells/cm2  is recommended.

Incubate cultures at 37°C. Subculture when cell concentration is between 3 X 105 and 4 X 105 cells/cm2 .

Subcultivation ratio: A subcultivation ratio of 1:4 to 1:10 is recommended.

Medium renewal: Every 2 to 3 days.

Reagents for cryopreservation

Complete growth medium supplemented with 10% (v/v) DMSO

Mycoplasma contamination

Not detected

Population doubling time

Approximately 16 hrs

【Organism物種來源】Animal  
【Tissue組織來源】
【Cell Type細胞類型】  
【Product Format產品狀態】 frozen/live culture
【Morphology細胞形態】  
【Culture Properties細胞特性】
【Biosafety Level生物安全級別】 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細胞源疾病】
【Age年齡】
【Gender性別】 Male/Female
【Storage Conditions保存條件】 liquid nitrogen vapor phase
【Karyotype染色體組型】
【Images細胞圖片】 Cell Micrograph
【Derivation衍生細胞】 -
【Clinical Data臨床數據】
【Comments其他描述】
【Complete Growth Medium完全培養基】 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傳代方法】
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凍存條件】
【Freeze Medium凍存培養基】Complete growth medium supplemented with 5% (v/v) DMSO
【Storage Temperature保存溫度】 Liquid nitrogen vapor phase
【Culture Conditions培養條件】
【Atmosphere培養環境】 Air, 95%; carbon dioxide (CO2), 5%
【Temperature培養溫度】 37°C
【STR Profile圖譜】
【Population Doubling Time倍增殖時間】
【References參考文獻】

【Supplier細胞供應廠家】BioVector NTCC Inc.
【Website網址】http://www.biovector.net