This document discusses the requirements and procedures for conducting in vitro studies according to Good Laboratory Practice (GLP). It outlines prerequisites like specialized infrastructure, training, and waste disposal arrangements. Prior requirements include justifying and characterizing the test system, and ensuring authenticity through identification, stability, functionality, and purity. Other responsibilities involve procedures for cryopreservation, maintaining sterility, and separating different studies and test systems. Proper documentation of all batch records, equipment maintenance, and cell origins must also be followed.

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