Using PGD to save affected children, Birth of healthy, HLA matched children

In some genetic diseases stem cell therapy can save lives. Thalassemia is the most prevalent disease in Iran. Another genetic disorder of interest was epidermolysis bullosa dystrophica (EB). In both cases a HLA matched child can help to save another sib’s life.PGD were carried out by selecting STR markers flanking either beta-globin gene or COL7A1 gene for beta-thalassemia and EB respectively. For HLA typing more than 52 STR markers flanking class 1 to class 2 genes as well as the centromere and telomere regions of the HLA region in chromosome 6 were used. These markers were used to construct haplotyping for the family and defective gene segregation.After confirmation of haplotype segregation, IVFs were carried out at different IVF centers collaborating with us particularly Abasn Hospital IVF center, which has shown to have the highest rate of success in producing viable fetuses and live born.A single blastomere was biopsied from each growing embryo. The blastomeres were put into each microtube containing lysis buffer. Each tube also contained primers for the specific gene (for sequencing), for STRs amplification and HLA typing. In each tube more than 25 sets of primers were amplified in two rounds of amplification. The second round contained labeled primers. For the second round a small amount of first round PCR products for 3-8 sets. The amplified products were run on sequencing machine.Haplotypings were carried out and the information was used to determine the fate of each embryo. HLA matched, unaffected embryos were determined and reported to the IVF center. So far 4 cases of HLA matched, disease PGD have resulted in births of healthy HLA matched children.We think PGD for HLA typing is feasible and can help families to have a healthy baby upon birth can make the family happy as well as hopeful to use small amount of his or her skin or bone marrow to save life of the affected sib.

کلیدواژه ها:

Embryo ، Genetic ، HLA ، IVF ، PCR