ADVANCED BIOMEDICAL TECHNOLOGIES

Second cycle degree in MEDICAL BIOTECHNOLOGIES

Campus: PADOVA

Language: English

Teaching period: First Semester

Lecturer: LEONARDO SALVIATI

Number of ECTS credits allocated: 8


Syllabus
Prerequisites: Advanced knowledge in virology, immunology and molecular biology. DNA structure and replication.
Examination methods: Written examination or oral presentation of recent scientific papers.
Course unit contents: ADVANCED BIOMEDICAL TECHNOLOGIES: The course will be comprised of 7 modules:

1) Genome structure: genes, pseudogens, SNPs, CNVs and repeated sequences. Bioinformatics databases. Relevance to molecular diagnosis

2) Mutations and mechanisms of dominance / recessivity. Allelic frequencies and Hardy-Weinberg's law. Database of genomic variants

3) Sanger sequencing and classic techniques for mutation detection.

4) Next generation sequencing in routine diagnostics: techniques, protocols, multigene panels, WES.

5) Interpretation of NGS data. Software and analysis strategies

6) Experimental models for the validation of variants

7) The problem of CNV. MLPA, RQ-PCR, CGH array and NGS

GENOME EDITING: Introduction to genome editing, Zinc Finger nucleases, Tale nucleases, CRISPR/Cas9. Problems related to the treatment of chronic viral infections, antiviral drugs resistance. Application of genome editing tools for the in vitro treatment of chronic viral diseases: HIV-1, HBV, HSV-1. Clinical trials for therapy of HIV-1.

CELL THERAPY: Stem cell research and biotechnology has great promise for the future of regenerative medicine. This has been the subject of great excitement and much progress has been made in understanding their mechanisms and unraveling their potential for therapeutic applications.
Limbal stem cell deficiency (LCSD) represents a major cause of visual impairment due to corneal opacification. It can occur unilaterally or bilaterally. The most frequent form is the unilateral LSCD caused by chemical or thermal injuries. When LSCD occurs, the neighboring conjunctival epithelium migrates over the cornea covering the corneal surface (corneal pannus) and therefore causing poor vision or even blindness. Conventional penetrating keratoplasty (PK) is not a therapeutic solution for pathologies or injuries leading to LSCD. The therapeutic use of limbal cultures for the permanent regeneration of corneal epithelium in patients with LSCD has been reported in many studies. According to the guidelines for good manufacturing practice (GMP), strictly regulated procedures and stringent quality control tests are now required to manipulate stem cells as “medicinal products” and make engraftment safer and eventually more successful.
This course describes techniques for optimal preparation of limbal stem cell grafts, including 1) a reliable impression cytology and imaging assays for the grading of LSCD, 2) culture methods that maintain high percentages of limbal stem cells, 3) the use of specific markers for the detection of corneal, conjunctival, and limbal stem cells, and 4) assays to evaluate the presence of contaminants, such as murine fibroblasts, endotoxins, mycoplasmae, and viral particles, in the cultured graft.
This course will take students on a journey into the stem cell biology, regenerative medicine and biotech revolution, revealing the modern scientific research behind stem cells and will provide the understanding of the various applications of this technology.