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Biology Chemistry DMPK Pharmacology Toxicology

Cell Cycle Analysis - PI Labeling

Service Description

Provider: Links Biosciences is a US company with laboratories in Hangzhou, China. The laboratory has been offering exploratory (non-GLP) pre-clinical services to US and Chinese biopharma since 2004.

Background: The intent of this service is to evaluate the effect of the Client’s test article on the cell cycle status of five diverse human tumor cell lines cultured under normoxic and hypoxic conditions. Flow cytometric interrogation of propidium iodide-labeled DNA is used to assign cell cycle stage. Cell cycle stage is ascribed according to DNA content; apoptotic (<2n), G0/G1 cells (2n) and S+G2 (>2n).

Service Details: Gaseous atmosphere: Control of the O2 tension in an atmosphere of 5% CO2 in air is achieved using N2. 20% O2 (normoxia) and 3% O2 (hypoxia) are the test atmospheres.

Service Details: Tumor cell lines: Enter 5 cell lines into the downloadable Sample Submission Form in the Sample Submission section. Fifty five (55) cell lines are available including: bladder cancer (5637); bone (SAOS-2, U2OS); breast (SKBR3); cervical (HeLa); esophageal (ECA-109); gastric (SW620, BGC-823, SGC-7901, HGC27, MGC803, AGS); glioma (U251); hepatic (HepG2, SMMC-7721, BEL-7402, H22{mouse}); kidney (786-0); leukemia (NB4, Molt-3, CEM, RS4;11, U937, P-388 {mouse}); lung (95D, Lewis {mouse}); mouth (KB); nasopharyngeal (CNE); neuroblastoma (SK-N-SH, CHP126, SH-SY5Y); ovarian (HO-8910, A2780); Drug or multi drug resistant cell lines include 2 breast, 3 leukemia and 1 mouth (KBR).

Service Details: Cell cycle status: The proportion of cells within each stage of the cell cycle is determined using propidium iodide (PI) labeling. Cells in logarithmic phase of proliferation are seeded in 6-well plates (1×105 cells/well) in 1 mL. For each cell line, cells are cultured with compound under conditions of normoxia and hypoxia. Cells are harvested at 6, 12, 24 and 48 h after addition of compound and washed with PBS. Cells are then incubated with PI and evaluated by flow cytometry. 15,000 events are analyzed to determine the proportion of cells that are apoptotic (<2n) or in G0/G1, S, or G2+M phases.

Regulatory Context – Study will be well controlled but will not be formally conducted in compliance with US GLPs.

Deliverable: Downloadable report containing all raw data and interpretation of results. This downloadable 2007 Leukemic cell line apoptosis report (479 KB) includes authors from Links Biosciences who contributed by performing this apoptosis assay (see figure 2b).

Price: This service evaluates one or more compounds in the test system described below. The service price under the add to cart button is to evaluate one (1) compound. Service price scales linearly so that the total cost for n compounds is this posted price multiplied by n. The service price for 1 or more compounds is calculated during the first step of the checkout process when the customer enters the number of compounds to be tested.

Sample Submission: Enter sample submission information and select service options in the Service Order Form that you can access once you add this item to your cart. The preferred compound form for submission is as a solid (powder). A solution may also be submitted. When submitting a solid, customer should provide dissolution instructions. Samples will be stored at approximately -80°C until testing.

How To Get Info

Click on the orange 'Request Info' button to request a custom quote, get bulk discount pricing, ask technical questions or request a meeting. Once you submit your request, you can communicate with the provider or exchange files by using Assay Depot’s microblogging system.

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How To Order

Click on the orange 'Add to Cart' button to access the Service Order Form, which contains technical questions and sample submission information. The form allows you to type in or paste in information or to select from a list of pre-defined choices. Complete and submit the Service Order Form to start the checkout process.

Once you submit your order, you can communicate with the provider or upload files by using Assay Depot’s microblogging system. The microblogging system allows you to stay in direct contact with the provider the entire time a research service is being carried out.


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