Cell Culture Laboratory
Head: Dr. Mario B. Guglielmi
The laboratory manages the Test Facility cell bank and associated activities. It conducts in vitro studies both compliant and non compliant with GLP standards. GLP-compliant studies mainly concerns medical device biocompatibility tests while non-GLP compliant studies mainly concerns Drug Discovery and ADME.
GLP-compliant Activities
- Biocompatibility
In accordance with the ISO 10993 standard (Biological Evaluation of Medical Devices), in vitro biocompatibility studies on medical devices and other materials and substances are conducted in order to assess the potential toxicity of medical devices and other materials and substances in contact with cells, tissues, or body fluids, simulating the interactions between the material used and the patient's tissues and the interaction between the device itself and cells, tissues, or body fluids.
This kind of activity includes:
- Cytotoxicity Tests
UNI EN ISO 10993-5 Standard: Tests for in vitro cytotoxicity
In vitro tests conducted in order to acquire, through specific tests (e.g. NRU and MTT), preliminary information on the general toxicity of the device, materials and chemical substances and to identify any biological damage caused by the substances released by the device through the observation of the effects that these produce on mammalian cells.
- Genotoxicity Tests (Mouse Lymphoma, Micronucleus)
UNI EN ISO 10993-3 Standard: Tests for genotoxicity, carcinogenicity and reproductive toxicity
In vitro gene mutation test on mammalian cells, widely used to determine the genotoxic potential of a substance, used in the screening of new molecules, in particular new candidate drugs and medical devices.
- Irritation, corrosion and sensitization tests
UNI EN IS10993-10 standard: Skin irritation and sensitization tests
UNI EN ISO 10993-23 standard: Test for irritation in vitro
OECD Guideline 439: In vitro skin Irritation
OECD Guideline 431: In vitro skin corrosion: human skin model test
OECD Guideline 442B: Local lymph node assay: BRDU-ELISA or FCM
In vitro and in vivo tests conducted to determine the acute irritant and/or corrosive effect of a chemical substance by evaluating the cytotoxic effect on RhE (Reconstructed Human Epidermis) models of reconstructed human epidermis (RhE) or the local inflammatory response to single or repeated applications of the substance under study at the skin level.
- Phototoxicity test (OECD Guideline 432: In vitro 3T3 NRU phototoxicity test).
The test allows to identify the in vitro phototoxic potential of a chemical substance activated by exposure to light. The test involves the use of 3T3 murine fibroblasts and is based on the comparison of the cytotoxicity of a chemical substance in the presence and in the absence of exposure to a non-cytotoxic dose of simulated sunlight. Chemical substances identified as positive by this test may be phototoxic in vivo after topical or systemic application to the skin and/or eyes.
Non GLP-compliant Activities
- Cell Banking
The availability of different cellular models and the possibility of choosing the most appropriate, makes their use convenient, reproducible and inexpensive in order to determine the biological activity and the toxicological profile of in vitro molecules and biological products. On the basis of specific experimental needs, it is possible to request:
- Culture, expansion, cryopreservation, tests for the identification of pathogens of over 100 stabilized tumor cell lines
- Isolation of primary cells from different tumor histotypes
- Creation of engineered cell lines that express reporter genes such as GFP or Luciferase
- Cytotoxicity and proliferation assays
Drug Discovery
In vitro cellular tests and screening platforms make possible to quickly characterize the compound under examination with reference to mechanism of action, cytotoxicity and ADME. The drug discovery activity includes the study of the cytotoxic, anti-proliferative, pro-apoptotic, differentiative effect of different classes of molecules and biological products on a wide range of human tumors. The different methods used (SRB, MTT, FACS analysis, Live Cell Imaging), applied in GLP-like conditions, make possible to characterize the potency, stability, mechanism of action, and toxicological profile of the molecules of interest, both individually and in combination with each other.
ADME Studies
- Metabolic Stability
Metabolic stability measures the stability of molecules in preclinical development using hepatocytes and hepatic microsomes of human or other animal species. This method allows to evaluate the metabolism of the molecule in vitro and to understand whether it is enzymatically stable or metabolized in the organism according to defined pathways.
- Intestinal Absorption
Intestinal absorption (permeability) is an important factor in order to evaluate the ability of a compound to cross the intestinal epithelium. This study is performed using the in vitro cellular model called CaCo2, allowing to classify the permeability (low, medium, high) of the compound according to the guidelines of the FDA’s Biopharmaceutics Classification System (BCS).