Conduct of in vitro studies in GLP setup

1. Conduct of In Vitro Studies in GLP
Setup
AB Pant
CSIR-Indian Institute of Toxicology Research
Lucknow-India
abpant@iitr.res.in, abpant@rediffmail.com

2. The Goal ???
To ensure the maximum reproducibility,
reliability, credibility and acceptance of
data produced
OECD No 14: Advisory Document (2004)
The Application of the Principles of GLP to in vitro Studies

3.  Studies which do not use multicellular organisms
as test systems
 Studies use microorganisms or material isolated
from organs as test systems
 In most cases, the OECD guidance on “Short-Term
Studies” applies
In Vitro Safety Studies: Definition

4. Prerequisites
 Specific infrastructure e.g. Aseptic work areas, CO2
Incubators, Microscopes, Cryocanes, etc.
 Specific training to study personnel to perform
highly critical procedures, with hazardous materials
 Specific arrangements for disposal of waste
materials to avoid risks of contaminating the
environment and other test systems

5.  Justification of the test system
 Characterisation of the test system
 Procurement details of cells
 Performance certificate from vender
 Details of positive, negative, untreated and/ or
vehicle controls
 Adequate performance of kits, their expiry, etc
Prior requirements

6.  Identification of cells: morphological
evaluation, genotypic and/or phenotypic
characteristics
 Stability and functional integrity of the system
in relation to its intended use
 Purity: freedom from biological contamination
Authenticity of the test system

7. Adequate training and knowledge of:
 Test procedures
 Aseptic condition and contamination risks
 Biohazard and chemicals
 Data interpretation

8. Identification of critical phases
Adequacy of batches of culture media, buffers,
antibiotics/ anti-mycotics, fetal bovine serum, etc.
Monitoring for potential cross contamination of
cells, mycoplasma, pathogens…
Cleaning & decontamination procedures
Use and maintenance of specialized equipment,
rooms, autoclaves, filters….
Other Responsibilities

9.  Cryopreservation and reconstitution of
cultures
 Processes that ensure sterility of supplies
used for cell and tissue cultures
 Maintenance of adequate separation between
different studies and test systems
Other Responsibilities

10. Points to be considered, but not be limited to:
Batch/ expiry dates of ingredients of culture media
Requisition/receipt forms for procurement of cells
Characterization/maintenance/ study records of cells
Contamination of undesired cells
Operation/ calibration/ maintenance procedures for equipments
Procedures ensuring the sterility
 Cryopreservation/ thawing/re-culturing of the cells
Separation between different studies & test systems
Implementation of SOPs of biosafety/ disaster management

11. • Ethical approval
• Species/strain of donor
• Tissue origin
• Sex of donor animal
• Age of fetuses or animals
used
• Health status
• Any special treatment
• Isolation procedure
• Cell type(s) isolated
• Date of isolation
• Name of user
Origin of cells and tissues
In case of cell line: passage number used

12. Thanks