Immortalized Human T Cells

Cat. No.
T0086
Unit
1x106 cells / 1.0 ml
Price
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Cat. No. T0086
Name Immortalized Human T Cells
Description

Immortalized Human T Cells were generated via lentiviral transduction of T cells isolated from a healthy donor.

Organism Human (H. sapiens)
Tissue Blood
Donor History Healthy donor, no other details disclosed
Growth Properties Suspension, round
Cell Type Immortalized Cells
Unit 1x106 cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
Intended 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.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions

PriGrow X Series Medium (TM0086) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. TM0086 has been optimized for T0086 and contains ROCK inhibitor. Additional IL-2 supplementation is not required. 

Note: To activate T cells, add 25 µl of Immunocult Human CD3/CD28 T Cell Activator (Stemcell #10971) per 106 cells, directly to the cell suspension, and incubate at 37°C and 5% CO2 for up to 3 days. For long-term expansion, subculture cells in TM0086 every 2-3 days and re-stimulate with the activator as required.


The post-thaw viability for T0086 is low. Do not activate the cells during the first 24 hours post-thaw. Activative cells after 24 hours and allow cells to grow undisturbed for next 3 days. Cells should form clumps once activated. Perform hemi-depletions of media and passage when required. 

Unpacking and Storage Instructions

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°C, 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°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.


Thawing Protocol

1. Thaw cells quickly in a 37°C 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

1. Simply add fresh complete media directly to the culture. Do not allow cell density to exceed 1x10⁶ cells/ml.

2. Alternatively, replace complete growth media by centrifugation and re-suspend the cell pellet in fresh complete media, and add appropriate aliquots of the cell suspension to new culture vessels, as desired.

3. Incubate the cells at the recommended conditions.

Cryopreservation

Cryopreservation Medium (TM024), or 50% complete growth media, 40% FBS,and 10% DMSO.

Immortalization Method

Transduction with Lenti-hTERT (LV616) lentivirus, Lenti-Rb siRNA (LV621) lentivirus, and a mixture of ten lentiviruses (ID1, ID2, ID3, Nanog, EZH2, Fos, SOX2, YAP1, LMO2 and KLF4)

Expression

CD3 and HLA-27 (via FACS)

STR Profiling

D5S818 : 11,12

D13S317 : 8,12

D7S820 : 10,12

D16S539 : 12,12

VWA : 16,17

TH01 : 6,9

AMEL : X,Y

TPOX : 8,11

CSF1PO : 11,12

D12S391 : 18,20

FGA : 19,21

D2S1338 : 18,20

D21S11 : 30,32

D18S51 : 12,19

D8S1179 : 15,16

D3S1358 : 14,18

D6S1043 : 15,20

PENTAE : 12,17

D19S433 : 13,14

PENTAD : 6,9

D1S1656 : 12,16

Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0086.
Warranty 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”.
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, 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 ordering. 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. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).

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 application(s). Please contact a technical service representative for more information.

4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and 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."
Application Research Use Only.
Material Citation If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0086
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