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Normal Primary Diploid Mortal NIH Swiss Mouse Embryonic Cells

SKU : BHC10902221

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T4138
Description	These primary normal fibroblast like NIH Swiss mouse cells (Cat. T4138) help define the starting normal genetic background of the NIH Swiss mouse diploid cell type.?These cells were prepared from fine minces of whole 17 to 19 days old NIH Swiss mouse embryos by the methods of Todaro and Green (ref 2) and grow well in the growth media used to grow all the R/J Model cells. This cell line is part of an eight cell R/J Model of Metastatic Progression designed using the NIH Swiss mice species to acquire isogenic cell types (T4138, T8981,T3317, T8144-T8148) that capture genetic/phenotypic stages in the progression to highly metastatic growth within this same species.?Compared to some other models, the metastasis related changes captured in this unique progression model are easier to discern above their less noisy isogenic background and have progressed within their native NIH Swiss in vivo micro-environments. This isogenic R/J Metastasis Cell Model was produced by an in vitro/in vitro?ix-step progression from mortal normal NIH Swiss mouse cells (Cat. T4138) that were used to produce immortal pre-cancer NIH/3T3 cells (Cat. T8981) that were transfected with the human HRAS oncogene to produce transformed cancer GhrasT-NIH/3T3 cells (Cat. T3317). Then beginning with a series of in vivo/in vitro progressive transfers of GhrasT-NIH/3T3 cells produced primary tumor T1-A cells (Cat. T8144) that produced local metastatic lesion T2-A cells (Cat. T8145) that simultaneously produced twin distant metastatic lung lesion T3-PA cells (Cat. T8146) with its twin distant metastatic liver lesion cells (Cat. T1847).?The?liver derived cells produced highly distant metastatic lung lesion cells (Cat T1848) which retained the human HRAS oncogene and they demonstrated a highly metastatic phenotype as they produced wide-spread simultaneous external and internal metastases in multiple sites in multiple mice when injected i.v. into nude NIH Swiss mice.Complete list of cells included in the R/J Metastasis Model: Cat.?8981??IH-3T3 CellsCat.?4138??ormal Primary Diploid Mortal NIH Swiss Mouse Embryonic Cells Cat.?3317??RAS Stably Expressing Transformed NIH/3T3 Cell Line (GhrasT-NIH/3T3)Cat.?8144? Primary Tumor Cell Line (T1-A ) Cat.?8145? Local Metastasis Tumor Cell Line (T2-A) Cat.?8146? Distant Pulmonary Metastasis Tumor Cell Line (T3-PA) Cat.?8147? Distant Hepatic Metastasis Tumor Cell Line (T3-HA) Cat.?8148? Highly Metastatic Distant Pulmonary Tumor Cell Line (T4-PA)
Cell Type	Primary Cells
Biosafety Level	2
Depositor	Grove City College (GCC)
Disclaimer	1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 2. 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 ?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. 3. 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). 4. 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. 5. 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. 6. 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."
Growth Properties	Adherent, fibroblast-like
Growth Conditions	PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.?riGrow III (TM003) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.Note: avoid excessive alkalinity in media; renew media twice weekly.
Tissue	Embryo
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	Mouse (M. musculus)
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.

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.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10902221	T4138	5x10^5 cells / 1.0 ml	-		Add to Quote

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