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Immortalized-Mouse-Natural-Killer-Cells-(KIL-C.2)

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Immortalized-Mouse-Natural-Killer-Cells-(KIL-C.2)

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Immortalized-Mouse-Natural-Killer-Cells-(KIL-C.2)

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	Print Version
BioSafety Level	II
Organism	Mouse
Source Organ	Blood
Growth Properties	Adherent
Morphology	Polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
Immortalization Method	Spontaneous immortalization and subcloning by limiting dilution
Description	A clonal derivative from the CD3^-NK1.1⁺NK cells developed from cocultures of OP-9S/LMJSN and post-5-FU bone marrow MNCs, the Immortalized Mouse Natural Killer Cells (KIL C.2) are dependent on IL-7 for proliferation and survival. Without culturing the cells in IL-7 the cells will undergo apoptosis within 24 hours. The cells are positive for CD43, CD51, CD94, and NKG2D surface markers which are comparable to the parental cells. If cultured with IL-2, the cells terminally differentiate exhibiting large azurophilic granules, cytoplasmic vacuoles, ruffled cell membranes, upregulation in granzyme B, and eventually die off in culture. It is recommended for use in studies relating to natural killer cell development.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is PriGrow V medium available from abm (TM015). To make the completed growth medium, add the following components to the base medium at a final concentration of 30% fetal bovine serum (TM999), 5x10^-5 M β-mercaptoethanol (CH045 ), 2mM L-glutamine (G275), 50 ng/ml recombinant mouse stem cell factor (Flt3L; Z200505), 25 ng/ml mIL-7 (R&D Systems; 407-ML-025/CF), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. Split ratio of 1:4 or 1:8 is recommended every 3 days. Footnote 1: cells grows best when conditioned medium of BHK/MKL cell line (secrets soluble rat SCF) is used at 10% (vol/vol) instead of pure SCF. Footnote 2: cells grows best when conditioned medium of J558 cell line (secrets murine IL-7) is used at 10% (vol/vol) instead of pure IL-7.
Quality Control	1) Flow cytometry analysis for cell surface markers; 2) Wright-Giemsa stain to visualize cells; 3) Western blost used to detect granzyme B
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. 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 strongly 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.