Media Preparation Core

Procedures and QC

Methods

The Media Lab at the Cleveland Clinic produces a variety of quality controlled tissue culture and bacteriological media, balanced salt solutions, tissue culture reagents, and buffers.

Media Lab glassware is soaked overnight in a sanitizing solution, washed in high pressure washers, baked at a high temperature of 180°C without caps, and then baked in a drying oven at 125°C for 8 hours using temperature sensitive sterilization monitors. Every bottle used goes through a final visual inspection before it is labeled and filled. Preparation of all products in the Media Lab begins with 0.2µm filtered high purity 18 MΩ-cm Super-Q water. Water is regularly tested for Endotoxins.

Filter

All Media Lab cell culture products are filtered through pharmaceutical grade double-layer 0.1µm filters from Pall Trincor offering complete retention of mycoplasma and removal of bacteria. Each filter is subjected to individual integrity testing by the manufacturer before it is certified and shipped to the Media Lab. In addition to the manufacturer’s testing, the Media Lab performs its own filter integrity test before and after filtration.

QC Broths

Prior to release, each batch of media is tested for sterility. A sample from each batch is mixed with nutrient broths. BHI (supports the growth of aerobic bacteria and yeast) SAB (supports fungal growth) and THIO (supports aerobic and anaerobic forms of bacteria). Testing is done in duplicate. After a 2-week test period, media is released for purchase.

Tissue Culture QC

Cell growth performance and colony forming efficiency tests are performed on each batch of tissue culture media. Mycoplasma testing is performed on each batch of tissue culture media and sterile solutions. Media Lab performs Endotoxin testing on all batches of media prepared.

Buffers and Microbiological Media

Sterility of autoclaved media and buffers is monitored with the use of biological indicators during the time of sterilization.

Carmel Burns
Department Manager, Media Prep Lab.
Email: burnsc@ccf.org