1. Eucaryotic Cell Bank Technology Transfer to the ‘BioMoBiL’ Centre 29 / 10 / Konrad Kleszczyński Jolanta Grzenkowicz – Wydra

2. Where from ?
ADVANCELL
A biotech company that offers customized research services on in vitro models (Toxicology, Oncology, Dermatology, Aging diseases)
Cell banking (156 cell lines)
Growing and expansion of new cell lines and their distribution
Cell lines quality control

3. Laboratory SOP Data Base Depositing Quality Control
Essential equipment to cell laboratory:
Flow Laminar Hood (Class II)
CO2 Incubator
Inverted Light Microscope
Fridge and Freezers: - 20 ÷ -30OC
- 80OC
Water Cleaning System (e.g. Millipore)
Centrifuges
Other equipment

4. Laboratory SOP Data Base Depositing Quality Control
Essential equipment for cell line depositing (Dewar’s containers):
Liquid nitrogen container
(3600 cryovials with the volume of 2 ml,
conatainer capacity: 116 L, evaporation: 0,6 L / 24 hrs)
Liquid nitrogen storage container
(container capacity: 34 L, evaporation: 0,25 L / 24 hrs)

5. Laboratory SOP Data Base Depositing Quality Control
Standard Operating Procedures
An SOP is a set of instructions to complete a job safely, with no adverse impact on the environment and in a way that maximizes operational and production requirements.
SOP performs:
Depositing of samples
Aparature: service, calibration, using
Chemicals: taking-up confirmation, labelling, protocols
All actions monitoring

6. Laboratory SOP Data Base Depositing Quality Control

7. Laboratory SOP Data Base Depositing Quality Control
DataBase organizes and menages with depositing cell lines
Access to the DataBase has just administrator who shows the place of the cryovial in the tank
Worker’s access to DataBase is limited to fill in the questionaire that is present in Intranet
All the actions are archived evereday

8. Laboratory SOP Data Base Depositing Quality Control

9. Laboratory SOP Data Base Depositing Quality Control
Services that are offered by Cell Line Collection
Cell line distribution
Safe deposit (short- and longlasting deposit)
Quality control
Mycoplasma eradication
Others

10. Laboratory SOP Data Base Depositing Quality Control
Depositing Organization of Bank Cell Line
Original Bank (OB)
up to 5 cryovials
Master Bank (MB)
cryovials
Distribution Bank (DB)
cryovials
106 cells / ml

11. Laboratory SOP Data Base Depositing Quality Control
Basic quality tests including: immunocytochemical test, test for presence of bacteria and mycoplasma detection
Mycoplasma detection is rapid, simple and sensitive method that uses PCR amplification with a commercial kit
Most of contaminations are caused by following mycoplasma species: M. arginini, M. fermentans, M. hyorhinis, M. orale, Acholeplasma laidlawii
Contaminated growing culture is treated by various antibiotics as tetracyclin derivatives (minocyclin, BM-cyclin)

12. Laboratory SOP Data Base Depositing Quality Control
C+ C- M
2% agarose gel electrophoresis of DNA amplification products
C+ positive control
C- negative control
M DNA molecular weight marker
1-7 examined samples
contaminated culture healthy culture with Mycoplasma
DNA staining by Hoechst 33258

13. Laboratory SOP Data Base Depositing Quality Control