Cell Culture Media Preparation Using MilliShot™ Single Dose Antibiotics

Section Overview

• Why Use Antibiotics?
• Important Points to Note Before You Start
• MilliShot™ Single Dose Antibiotics Protocol
• Related Products

Why Use Antibiotics?

Antibiotics are crucial reagents for cell culture researchers to prevent contamination of cultures from pathogens and to minimize the loss of reagents and cells. Bacterial and fungal contaminants can induce physiological and morphological changes in cultured cells, affecting experimental outputs. Antibiotics can also be used to select for genetically modified cells during experimental assays. Therefore, antibiotics, in addition to aseptic conditions, are vital for reproducible tissue culture data.

While the most frequently used antibiotics to prevent contamination are Penicillin-Streptomycin or Antibiotic-Antimycotic solution, some scientists may add Amphotericin B or Gentamicin to the medium. Here we describe the protocol for preparing culture media using our MilliShot™ single dose antibiotics. MilliShot™ single dose antibiotic solutions are cell culture tested, ready-to-use antibiotics, conveniently packaged in one-time use vials, saving time spent preparing cell culture media.

Important Points to Note Before You Start

• All steps of media preparation and cell culture must be performed in an aseptic laminar flow hood.
• Confirm the right media and supplements for your cell line. Download the ECACC handbook for all information connected to the cell line of your interest.
• Bring your liquid media, FBS and supplements to 37°C before you start.

MilliShot™ Single Dose Antibiotics Protocol

Step 1

Remove MilliShot™ single dose antibiotics vial from the freezer.

Step 2

Supplement media of choice with MilliShot™ single dose antibiotics vial.

Step 3

Sterile filter your media using Stericup^® filter devices.

Step 4

Culture cells according to cell type.

All the steps are performed in laminar flow chamber under aseptic conditions.

1. Open a 500mL bottle of medium suitable for your cell line, such as DMEM, RPMI, or MEM with 2mM glutamine.
2. Add 50mL of FBS of your choice to prepare medium with 10% FBS. For cell lines that need 5% FBS supplementation, add 25mL of FBS of your choice.
3. If your medium does not have added glutamine, add 5mL of 2mM glutamine. Add appropriate concentration of any other growth supplements that your cell line may require for growth.
4. Open and add the entire contents (5mL) of one vial of MilliShot™ single dose Penicillin-Streptomycin/Antibiotic-Antimycotic solution/Amphotericin B/Gentamicin to the medium. You can directly pour or use a pipette to add the vial contents to the medium.
5. Filter the prepared complete medium using Stericup^® filtration devices to obtain sterile complete medium to culture your cell lines.
6. Culture your cells as required for your cell type and experimental outcome.