Maintenance medium change

1. Scope

·       This document contains procedure to change the tubes and flasks with maintenance medium.

2. Definition

·       RD-     Human Rabdomyosarcoma cell line sensitive to Polio virus and many other Enteroviruses.

·       L₂₀B -    Cell line derived from mouse L-cells transfected with polio receptors that will support the growth of Polioviruses and very few other enteroviruses.

·       MEM- Eagles Minimum Essential Medium.

3. Material

·       10 ml pipette

·       Pippetaid

·       Prepared tubes and flasks

·       2% maintenance medium

4. Procedure

·       When monolayer is nearly confluent (2-3days), change tubes to maintenance medium.

·       On third day check the tubes under microscope.

·       After that take one by one rack of tubes (L20B and RD) in Biosafety cabinet.

·       Decant all tubes one by one.

·       After that add maintenance medium with 10 ml pipette, add 1ml medium one by one.

·       These tubes used for inoculation.

·       In case of flask, change the medium on 4^th or 5^th day.

5. Safety conditions

·       Never handle different cell culture at the same time.

·       Be careful to prevent microbial contamination to cell culture and contamination of working environment with cell culture material.

                         

6. Documentation  

• Log Book of change with 2% MEM medium PR-42

7.    References used to draw SOP

·       WHO Polio Laboratory Manual.