PGDIS HANDS-ON WORKSHOP ON PGD FOR GENETIC AND CHROMOSOMAL DISORDERS BY TROPHECTODERM BIOPSY

Thursday, April 11, 2019

09:00 - 10:05   PGD for Single Gene and Chromosomal Disorders by Blastomere and Trophectoderm Biopsy - Theoretical Session 1

08:30 - 09:00   Registration

09:00 - 09:05   Opening of Workshop, Semra KAHRAMAN, Memorial Hospital ART and Genetics Center, Istanbul, Turkey

09:05 - 09:35   Practical PGT, Anver KULIEV, Reproductive Genetics Institute, Chicago, USA

09:35 -10:05    Advantages and Disadvantages of Trophectoderm Biopsy, Hakan YELKE, Memorial Hospital ART and Genetics Center, Istanbul, Turkey

10:05 -10.35    DISCUSSION & COFFEE BREAK

10:35 -13:10    Hands-on Applications of Biopsy Procedures under Supervision of Trainers Participants will be divided into 3 groups, up to 5 in each for practical sessions.

• Zona Drilling Methods
• Blastomere Biopsy Techniques
• Trophectoderm Biopsy Techniques

13:10 - 14:20   LUNCH & DISCUSSION

14:20 -18:30    Each group will apply all the demonstrated techniques

Friday, April 12, 2019

09:00 - 10:05   PGD for Single Gene and Chromosomal Disorders by Blastomere and Trophectoderm Biopsy- Theoretical Session II

09:00 - 09:20   Clinical Outcome in PGT-A and What to Do with Mosaic Embryos?                  Semra KAHRAMAN, Memorial Hospital ART and Genetics Center, Istanbul, Turkey

09:20 - 09:40   PGT for Aneuploidy (PGT-A) and Structural Rearrangements (PGT-SR)-From FISH to

                        Next Generation Technologies, Anver KULIEV, Reproductive Genetics Institute, Chicago, USA

09:40 - 10:00   Molecular Techniques and Next Generation Technologies in PGT: Testing for Monogenic Disorders (PGT-M), HLA Typing (PGT-HLA), and 24 Chromosome Aneuploidy (PGT-A) by Next Generation Sequencing (NGS), Don LEIGH, Sydney, Australia

10:05 -10:30    DISCUSSION & COFFEE BREAK

10:30 -13:30    Hands-on applications of trophectoderm biopsy by participants under supervision of trainers

13:30 -14:30    LUNCH & DISCUSSION

14:30 -18:30        Hands-on Applications of FISH and PCR techniques under supervision of trainers

• Preparation, Spreading and Fixation of Blastomere and Trophectoderm Cells
• In Situ Hybridization with Different Chromosome Specific Probes
• Analysis and Scoring of FISH Signals
• 1st and 2nd Round PCR for PGD of Single Gene Disorders and HLA Typing
• Testing for Next Generation sequencing

Participants will be divided into 3 groups, up to 5 in each for practical sessions according to their interests.

Saturday, April 13, 2019

09:00 - 13:00   Interpreting FISH, NGS and PCR Results

09:00 - 12:00   Analysis and Scoring of FISH Signals, and NGS results, Don LEIGH, Syndey, Australia and Murat CETINKAYA, Memorial Hospital ART and Genetics Center, Istanbul, Turkey.

Analysis and Interpretation of PCR Results, Don LEIGH, Syndey, Australia and Murat CETINKAYA, Memorial Hospital ART and Genetics Center, Istanbul, Turkey

12:00 -13:00    GENERAL DISCUSSION & CLOSING

Practical Stations

Station 1:        Blastomere Biopsy Techniques;

• Zona hatchings by enzymatic, mechanical and laser applications

Station 2:        Trophectoderm Biopsy Techniques;

Station 3:        FISH Techniques;

• Preparation, spreading and fixation of Blastomere and Trophectoderm Cells
• In situ hybridization of chromosomes with different probes
• Analysis and Scoring of FISH signals

Station 4:        NGS Techniques;

• Extraction, amplification and barcoding of blastomere and trophectoderm cells
• Pooling, purification and quantification of the samples
• Emulsion PCR and enrichment
• Chip loading and analysis of NGS results

Station 5:        PCR Techniques;

• Lysis of blastomere and trophectoderm cells
• First and second round PCR for PGD of single gene disorders and HLA matching Mutation Analysis and Restriction Enzyme Digestion
• Analysis and interpretation of PCR results

• These applications will be done simultaneously in 4 stations and each participant will attend to each station and perform all demonstrated techniques