Cell Authentication Testing

AUTH10: 10 markers in the STR profiling analysis

AUTH16: 16 markers in the STR profiling analysis

AUTH10P: same as AUTH10 plus Profile Search

AUTH16P: same as AUTH16 plus Profile Search

Background

Cross-contamination and misidentification of cell lines are unfortunately very common within the research community. Many contaminated cell lines were overgrown by the contaminating cell line during establishment and so authentic stocks probably do not exist; in those cases all work has been performed on the contaminant, which may come from a different species, tissue or cell type.

To let researcher check the true identity of their cell lines to have a peace of mind- a requirement increasingly imposed by scientific journals and funding agencies - AddexBio offers a cell line authentication service.

General recommendations are to authenticate:

1. When a new line is established or acquired to determine an identity for the cell line.

2. Before freezing.

3. When the culture is actively growing for 2-3 months.

4. When the performance of the cell line is not consistent or results are unexpected.

We offer 2 levels of service, Basic Service and Basic Service with Profile Search.

The Basic Service includes a authentication report for each line submitted that is suitable for proof of cell line authentification which may be required by some journals and grant funding agencies. The results are used to check for cross contamination, cell line mix-ups and percent match between lines or to establish the identity of a new cell line.

Basic Service with Profile Search includes the above, together with cross referencing against repositories ATCC, DSMZ, and JCRB databases to check the authenticity of your cell line.

Service Provided

We provide STR profiling analysis for cell lines of human origin by showing different numbers of markers.

AUTH10: GenePrint 10 System (Promega), Markers included in this analysis are amelogenin, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, THO1, TPOX, and vWA.

Or

AUTH16: PowerPlex 15HS (Promega), Markers included in the kit are amelogenin, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, THO1, TPOX, vWA, D18S51, D3S1358, FGA, D8S1179, Penta E, and Penta D.

Ordering

We require that customers contact our customer support to submit an order form to use our Cell Line Authentication Service before sending samples to us.

Procedures for Submitting Cell Cultures for Cell Line Authentication Service

1. Samples should be submitted from growing culture (please see below for detailed cell culture preparation). We accept only BSL-1 and BSL-2 samples for testing.

2. Prior to sending the cells for testing, please contact customer support and submit an order form either by email at customersupport@addexbio.com or by fax at 858-538-8847, so that we will be receiving the cells.

3. If you are submitting more than one cell culture sample, place samples for each culture in separate zipper-locked or sealed bags. This will protect the integrity of the other sample(s) if one of them leaks.

4. Fill out the order form that you received from customer support; make sure you include your reference number. Pack cell culture sample(s) at room temperature in a sturdy box to prevent breakage. Each flask/tube should be wrapped in plastic bubble wrap or comparable packing material to prevent breakage or leakage. Additional packing material should be used for protection in shipping. Do not use vented caps as they may leak during shipping.

5. Send materials at room temperature by courier, such as FEDEX or UPS (maximum 2-day delivery for live cells, overnight for frozen vials). Materials must arrive at AddexBio by noon on Monday through Thursday of the week. Please include signed order form with sample(s).

Cell Culture Preparation for Cell Line Authentication Service

3. Prepare samples(s) following one of the procedures based on cell type:

For Preparation with Adherent Cells

1. Aseptically seed a sterile T-25 with approximately 5x10⁴ cells in 10 mL of medium and grow for 2 population doublings (about 48 hours).

2. Without replacing the current culture medium, aseptically fill the flask completely with fresh culture medium. Seal the lid of the closed flask with Parafilm sealer.

For Preparation with Suspension Cells

Aseptically place approximately 2x10⁵ cells in a sterile container topped up with medium, container can be either a small plastic tube with a screw-cap closure or a T-25 flask. Seal the lid of the closed container with Parafilm sealer.

For Frozen vial

Keep frozen vial(s) in a foam package that has sufficient dry ice for the delivery.

Samples received that show signs of leaking or obvious bacterial/fungal contamination will not be tested.

We do not accept culture samples(s) containing HIV, virulent microbials (fungi, bacteria or viruses), or other BSL 3 or 4 agents.