Procedure Outline: Purchase Cell Line from Repository -----> Log Lot and Passage Received -----> Thaw Vial Received -----> Expand to Freeze ~5 vials -----> Validate/Mycoplasm Test -----> Thaw Frozen Vial to Test Freeze Batch -----> Compare Growth in Recommended Media Compared to advanced RPMI with 4% certified FBS and 1x Glutamax (when possible switch to aR4) -----> Expand selected Media Culture to Freeze 11-15 vials -----> Validate/Mycoplasm Test -----> Thaw Frozen Vial to Test Freeze Batch -----> Expand to Collect Sufficient Cells for Funded Studies and Potential Future Studies -----> Validate/Mycoplasm Test. * Passage numbers maintained through entire process. If provided from repository the passage is additive if not starts from passage 0 * Cell lines will be validated using the CNV fingerprint method (Keats et al. Submitted) * Mycoplasm screen is conducted using MycoAlert Testing Kit from Lonza *NCA* - Not Currently Available for Purchase |
