JavaScript is currently not supported/disabled by this browser. Please enable JavaScript for full functionality.

Call Us 1 - 866 - 986 - 9598
orders@biohippo.com

Sign Up

Login
- My Order
- Return Requests
- Track my Order

Home / Cells / Cell Lines

Immortalized Human Diploid Fibroblasts - hTERT

SKU : BHC10900542

Contact us to order

Tel

+1 866.986.9598

Email

orders@biohippo.com

Get a Quote

Request More Infos

Applied Biological Materials (abm) Inc.

view supplier details

Order Now

Credit card payments now incur a 3% fee.

Contact us to order

Tel

+1 866.986.9598

Email

orders@biohippo.com

Get a Quote

Ask Merchant

Request More Infos

Sample Request

Mfr.No.	T1013
Description	Real Time PCR was used to quantify hTERT gene expression in immortalized cell line
Cell Type	Immortalized Cells
Biosafety Level	2
Disclaimer	1. All test parameters provided in the CoA are conducted using abm's standardized culture system and The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s). 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period".
Donor History	Female, 3 months, Caucasian
Expression Level	DAT, VMAT-2, TH, alpha-SYN, beta-III tubulin
Growth Properties	Adherent, fibroblast
Growth Conditions	Use of PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow III (TM001) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?
Quantity	1x106 cells / 1.0 ml
Tissue	Lung
Morphology	Flattened\|Dendritic processes
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Human (H. sapiens)
Population Doubling	30 - 40 hours
Species	Human (H. sapiens)
Propagation	Grow the cells in culture vessel pre-coated with 50 mug/ml poly-L-ornithine (PLO) (TM062) and 1 mug/ml fibronectin (EMD Millipore; Cat. FC010) in H2O for at least 3 hours at 37°C. Do not grow these cells in culture vessels with surface areas equal to less than 12.5 cm2. These cells do not grow in 6-well, 24-well, 48-well, or 96-well plates.The base medium for this cell line is Advanced DMEM/F12 (Gibco;12634010). To make the complete growth medium, add the following components to the base medium: N2 supplement (ThermoFisher Scientific) to a final concentration of 1X, L-glutamine (G275) to a final concentration of 2 mM, and Recombinant Human FGF2 (Z101455) to a final concentration of 40 ng/ml.Change media every 2-3 days. Do not let media colour change to orange-yellow.The cells will form round clumps instead of a monolayer when stressed. It is recommended to subculture when cells are grown to a cell density of 50-60%.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.To subculture the cells, use 0.25% Trypsin-EDTA (TM051). After adding trypsin, incubate the cells at room temperature for 2-3 minutes and agitate the culture vessel until 90% of the cells have detached. Immediately neutralize the trypsin using Trypsin Neutralizing Solution (Lonza, Cat. CC-5002). Add Trypsin Neutralizing Solution twice the volume of trypsin used. Centrifuge at 200x g for 2-3 minutes. Aspirate supernatant and resuspend cells in complete media. Plate cells into pre-warmed PLO-fibronectin-coated vessels at 37.0°C. Avoid subculturing if the cells appear stressed. Note: If leaving the cells over the weekend (or more than 2 days), make sure to do a high split ratio (1:4 to 1:5).To differentiate the cells into neurons, change the medium to differentiation medium after the cells have grown to a density of 40-50%.To make complete differentiation medium, add the following components to Advanced DMEM/F12 (Gibco;12634010): N2 supplement to a final concentration of 1X, L-glutamine (G275) to a final concentration of 2 mM, dbcAMP to a final concentration of 1 mM, tetracycline to a final concentration of 1 mug/ml, and Recombinant Human GDNF (Z101055) to a final concentration of 2 ng/ml. Allow the cells to grow in differentiation medium for 4-6 days before testing for neuron specific markers.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control	mRNA and protein expression levels of various markers pre and post differentiation are verified by RT-PCR and western blotting.

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol	Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.

Have you used this product?

Rate it now..!!

Login to Review

SKU	Supplier No.	Size	Your price	Quantity
BHC10900542	T1013	1x10^6 cells / 1.0 ml	-		Add to Quote

ebiohippo.com provides biomedical researchers with a one-stop informatic shopping experience by transforming biotechnology products’ data into a ready-to-analyze state. We collect and unify suppliers’ data on a single platform to reduce the time of searching, uncertainty, and cost of scientific experiments.

© 2024 Biohippo. All rights reserved.