MDA-MB-435S

This cell line was originally described as a spindle shaped variant of the parental MDA-MB-435 strain isolated in 1976 by R. Cailleau, et al. from the pleural effusion of a 31 year old female with metastatic, ductal adenocarcinoma of the breast. 

However, recent studies have generated questions about the origin of the parent cell line, MDA-MB-435, and by extension HTB-129. Gene expression analysis of the cells produced microarrays in which MDA-MB-435 clustered with cell lines of melanoma origin.

This cell line was originally described as a spindle shaped variant of the parental MDA-MB-435 strain isolated in 1976 by R. Cailleau, et al. from the pleural effusion of a 31 year old female with metastatic, ductal adenocarcinoma of the breast. 

However, recent studies have generated questions about the origin of the parent cell line, MDA-MB-435, and by extension HTB-129. Gene expression analysis of the cells produced microarrays in which MDA-MB-435 clustered with cell lines of melanoma origin instead of breast [PubMed ID: 10700174, PubMed ID: 15150101, PubMed ID: 15679052]. 

Additional studies have since corroborated a melanocyte origin of MDA-MB-435, to which ATCC has responded by pursuing its own investigation into the identity of this cell line. The cell line to which MDA-MB-435 is reported to have been cross-contaminated with is the M14 melanoma line [PubMed ID: 12354931 and PubMed ID: 17004106].

1. Check all containers for leakage or breakage.
2. 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.

• 0.01mg/ml bovine insulin
• 0.01mg/ml glutathione
• fetal bovine serum to a final concentration of 10%

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.

1. 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).
2. 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.
3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to7 minutes.
4. 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 culture flask. 
5. Incubate the culture at 37°C in a suitable incubator. 

The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO₂ and air mixture is detrimental to cells when using this medium for cultivation

Remove medium, add fresh 0.25% trypsin - 0.53 mM EDTA, rinse and remove. Place flask at room temperature (or incubated at 37°C) for approximately 10 minutes or until the cells detach. Add fresh medium, aspirate and dispense into new flasks.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

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