presentation instabul +videos 1 6-2010 (1)

MARINOS TSIRIGOTISMARINOS TSIRIGOTISMD.FRCOGMD.FRCOGDIRECTOR “AKESO” FERTILITY CENTERDIRECTOR “AKESO” FERTILITY CENTER

22ndnd international Meeting international Meeting ““New Perspectives on ovulation induction”New Perspectives on ovulation induction”

3-4 June 20103-4 June 2010IstanbulIstanbulTurkeyTurkey

Is There any Indication for PGD?Is There any Indication for PGD?

Types of Preimplantation Genetic TestingTypes of Preimplantation Genetic Testing


Preimplantation Genetic DiagnosisPreimplantation Genetic Diagnosis -PGH-PGH

Preimplantation Genetic ScreeningPreimplantation Genetic Screening

Marinos Tsirigotis, MD.FRCOG, Director “AKESO” Fertility CenterMarinos Tsirigotis, MD.FRCOG, Director “AKESO” Fertility Center

What is Preimplantation genetic Testing?What is Preimplantation genetic Testing?


The procedure which involves the removal of one or more The procedure which involves the removal of one or more nuclei from oocytes (polar bodies) or embryos nuclei from oocytes (polar bodies) or embryos (blastomeres or trophectoderm cells) to test for mutations(blastomeres or trophectoderm cells) to test for mutations in gene sequence or aneuploidy before transfer.in gene sequence or aneuploidy before transfer.


What is Preimplantation Genetic Diagnosis ? What is Preimplantation Genetic Diagnosis ?


When one or both genetic parents carry a gene mutation When one or both genetic parents carry a gene mutation or a balanced chromosomal rearrangement and testing is or a balanced chromosomal rearrangement and testing is performed to determine whether that specific mutation or performed to determine whether that specific mutation or an unbalanced chromosomal complement has been an unbalanced chromosomal complement has been transitted to the oocyte or embryo.transitted to the oocyte or embryo.

PGDPGD


Indications forIndications for


■ ■ Autosomal dominant disorders (50% risk)Autosomal dominant disorders (50% risk)

■■ Autosomal recessive disorders (25%)Autosomal recessive disorders (25%)

■■ Female carriers of x-linked disordersFemale carriers of x-linked disorders (25% of female, & ½ of male embryos)(25% of female, & ½ of male embryos)

■■ Previous affected child with a de-novo mutationPrevious affected child with a de-novo mutation (ie: Tuberous Sclerosis)(ie: Tuberous Sclerosis)

■■ Human leukocyte antigen (HLA) matchingHuman leukocyte antigen (HLA) matching (b-thalassaemia, fanconi anaemia, ALL, DBA etc.)(b-thalassaemia, fanconi anaemia, ALL, DBA etc.)

■■ Carriers of a balanced chromosomal translocation, inversion Carriers of a balanced chromosomal translocation, inversion or other structural chromosomal rearrangement or other structural chromosomal rearrangement (ie: markers)(ie: markers)

■■ Cancer predispositionCancer predisposition

PGDPGD



Autosomal dominant conditions Autosomal dominant conditions (most frequently diagnosed)(most frequently diagnosed)

Myotonic dystrophyMyotonic dystrophy Huntington's diseaseHuntington's disease Charcot Marie toothCharcot Marie tooth Adenomatous polyposis coliAdenomatous polyposis coli Von Hippel-LindauVon Hippel-Lindau NeurofibromatosisNeurofibromatosis Marfan syndromeMarfan syndrome Breast cancerBreast cancer Tuberous sclerosisTuberous sclerosis Osteogenesis imperfectaOsteogenesis imperfecta RetinoblastomaRetinoblastoma



Autosomal recessive conditionsAutosomal recessive conditions

Cystic fibrosisCystic fibrosis β-thalassemiaβ-thalassemia Spinal muscular atrophySpinal muscular atrophy Sickle cell diseaseSickle cell disease 21-hydroxylase deficiency21-hydroxylase deficiency Tay-Sachs diseaseTay-Sachs disease Gaucher diseaseGaucher disease Epidermolysis bullosaEpidermolysis bullosa Fanconi anemiaFanconi anemia



x-linked diseasesx-linked diseases

Sex-linked recessiveSex-linked recessive (50% of males will be affected)(50% of males will be affected)

● Haemophilia AHaemophilia A● Becker muscular dystrophyBecker muscular dystrophy● Retinitis pigmentosaRetinitis pigmentosa● AdrenoleukodystrophyAdrenoleukodystrophy● Lesch Nyhan syndromeLesch Nyhan syndrome● Spinal bulbar muscular atrophySpinal bulbar muscular atrophy Sex-linked dominantSex-linked dominant (50% of males will be affected,(50% of males will be affected,females may be affectefemales may be affected)d)

● Duchenne muscular dystrophyDuchenne muscular dystrophy● Alport syndromeAlport syndrome● Fragile X syndromeFragile X syndrome● Incontinentia pigmentiIncontinentia pigmenti


Preimplantation genetic screening (PGS)Preimplantation genetic screening (PGS)Chromosomal aneuploidy screeningChromosomal aneuploidy screening


When genetic parents are known or presumed to be When genetic parents are known or presumed to be chromosomally normal and their embryos are screened chromosomally normal and their embryos are screened for aneuploidy.for aneuploidy.


Indications for PGSIndications for PGS


■ ■ Advanced Maternal Age (AMA)Advanced Maternal Age (AMA)

■■ Recurrent early Pregnancy Loss (RPL)Recurrent early Pregnancy Loss (RPL) (particularly with chromosomally abnormal foetuses)(particularly with chromosomally abnormal foetuses)

■■ Repeated IVF (implantation) failure (RIF)Repeated IVF (implantation) failure (RIF)

■■ Severe male infertilitySevere male infertility

■■ Patients with sex chromosomes aneuploidyPatients with sex chromosomes aneuploidy

■■ Sex selectionSex selection

■■ Single embryo transferSingle embryo transfer

■■ On request On request (patients without known genetic disorders)(patients without known genetic disorders)


How is PGD/PGS performed?How is PGD/PGS performed?


■ ■ Polar Body biopsyPolar Body biopsy

■■ BlastomereBlastomere

■■ Trophectoderm biopsyTrophectoderm biopsy


Why Polar Body biopsy?Why Polar Body biopsy?


Polar body is found outside the ooplasm or embryo, so Polar body is found outside the ooplasm or embryo, so the embryo is not disturbed the embryo is not disturbed

In some countries, social and ethnical reasons do not In some countries, social and ethnical reasons do not allow for embryo biopsyallow for embryo biopsy

Blastomere testing for aneuplodies has an important Blastomere testing for aneuplodies has an important limitation due to the extremely high rate of mosaicism limitation due to the extremely high rate of mosaicism at the cleavage stageat the cleavage stage


Indications for *Polar Body biopsy?Indications for *Polar Body biopsy?


Maternally Derived Mendelian diseaseMaternally Derived Mendelian disease (single-gene disorders)(single-gene disorders)

■■ Mother – carriers of chromosomal abnormalitiesMother – carriers of chromosomal abnormalities

■■ Aneuploidy screening due to chromosomal Aneuploidy screening due to chromosomal pathology in AMApathology in AMA

** Only maternally derived disorders can be detected Only maternally derived disorders can be detected


Methods of Genetic AnalysisMethods of Genetic Analysis


■■ PCRPCR (Polymerase Chain Reaction)(Polymerase Chain Reaction)


for specific gene mutation disorderfor specific gene mutation disorder

■■ FISH FISH (Fluorescence in situ hybridization)(Fluorescence in situ hybridization) for aneuploidy screening & carriers of chromosomal abnormalitiesfor aneuploidy screening & carriers of chromosomal abnormalities

■■ CGH CGH (Comparative Genomic Hybridization)(Comparative Genomic Hybridization)

- array CGH- array CGH - SNP array- SNP array

Limitations ofLimitations of PGDPGD


■■ Relatively short interval of time available for Relatively short interval of time available for analysisanalysis

■■ Only one or two cells can be analysedOnly one or two cells can be analysed

■■ In Polymerase Chain Reaction based methodsIn Polymerase Chain Reaction based methods misdiagnosis or non-diagnosis from:misdiagnosis or non-diagnosis from:

- anucleate cell- anucleate cell - failure of amplification of targeted DNA segment- failure of amplification of targeted DNA segment - external contamination- external contamination - allele drop-out- allele drop-out - partial amplification- partial amplification

■■ Estimated risk of transferring an affected embryo Estimated risk of transferring an affected embryo mistakenly identified as normal by PGD is mistakenly identified as normal by PGD is

• 2% for recessive disorders2% for recessive disorders• 7-8% on average for dominant disorders7-8% on average for dominant disorders


Limitations ofLimitations of PGS PGS


■■ FISH allows evaluation of fewer than half of the 23 FISH allows evaluation of fewer than half of the 23 chromosomes pairs (X,Y, 1, 13,16,17,18,21,22)chromosomes pairs (X,Y, 1, 13,16,17,18,21,22)

■■ Up to 25% of aneuploid embryos will be found Up to 25% of aneuploid embryos will be found normal by FISH because the abnormal chromosomesnormal by FISH because the abnormal chromosomes are not among those included in the analysis are not among those included in the analysis

■■ Approximately 10% of cells removed for screening yield Approximately 10% of cells removed for screening yield no resultsno results

■■ Splitting or diffused signals due to overlapping or Splitting or diffused signals due to overlapping or misorientation of chromosomesmisorientation of chromosomes



■■ Failure to Hydridize Failure to Hydridize

■■ A mosaic embryo can be better identified only if two A mosaic embryo can be better identified only if two or more cells are removed and analyzed. Exclusion of or more cells are removed and analyzed. Exclusion of pathology requires all of the cells to be tested, pathology requires all of the cells to be tested, destroying the embryo destroying the embryo

■■ Even if two cells are removed from each embryo Even if two cells are removed from each embryo we can still get discordant results. we can still get discordant results. Practically half of the embryos will be euploid after all Practically half of the embryos will be euploid after all cells have been analyzedcells have been analyzed



■■ Up to 50% of those embryos identified as aneuploid Up to 50% of those embryos identified as aneuploid at the cleavage stage will “self correct” at blastocyst at the cleavage stage will “self correct” at blastocyst stage stage

■ ■ AAbnormal results from FISH analysis of a single bnormal results from FISH analysis of a single blastomere on day 3 embryo do not necessarily blastomere on day 3 embryo do not necessarily indicate that the embryo is abnormalindicate that the embryo is abnormal

■■ ET at blastocyst stage ET at blastocyst stage


Limitations of Limitations of PGSPGS


Comperative Genomic Hybridization (CGH)Comperative Genomic Hybridization (CGH) or Chromosomal Microarray Analysis (CMA)or Chromosomal Microarray Analysis (CMA)

■■ CGH will detect only unbalanced chromosomal changesCGH will detect only unbalanced chromosomal changes

■■ Structural chromosome aberrations (reciprocal, inversions, etc) Structural chromosome aberrations (reciprocal, inversions, etc) can not be detectedcan not be detected

■■ Array CGH overcomes many of these limitationsArray CGH overcomes many of these limitations

■■ However, both CGH & array CGH do not provide information However, both CGH & array CGH do not provide information as to the ploidy. Balanced DNA content ie: tetraploid clone as to the ploidy. Balanced DNA content ie: tetraploid clone will appear normal will appear normal


Limitations ofLimitations of PGD PGD && PGS PGS


■■ Embryologist experienceEmbryologist experience

■■ Biopsy techniqueBiopsy technique

■■ Spreading Technique (ie: air bubble!)Spreading Technique (ie: air bubble!)

■■ Number of oocytes – embryos (ie: AMA, OHSS)Number of oocytes – embryos (ie: AMA, OHSS)

■■ Embryo morphologyEmbryo morphology


Does biopsy affect the embryo?Does biopsy affect the embryo?


■■ Theoretically removal of 1 or 2 blastomeres should not decrease Theoretically removal of 1 or 2 blastomeres should not decrease pregnancy ratespregnancy rates

■■ However, there have been reports whereby pregnancy rates However, there have been reports whereby pregnancy rates have been lower after embryo biopsy compared to have been lower after embryo biopsy compared to conventional IVFconventional IVF

■■ This may be due to the reduced implantation potential of This may be due to the reduced implantation potential of embryos undergoing biopsyembryos undergoing biopsy

■■ Biopsy technique, experience & culture conditions important Biopsy technique, experience & culture conditions important co-factorsco-factors



■■ Indications “per-se” PRsIndications “per-se” PRs

■■ Early growth and risk of congenital malformations not Early growth and risk of congenital malformations not different to other ART children different to other ART children (ESHRE task force of PGD)(ESHRE task force of PGD)

■■ However, PGD may pose neurological risksHowever, PGD may pose neurological risks (memory decline and increased weight in adulthood in mice)(memory decline and increased weight in adulthood in mice)


Clinical applications of Clinical applications of PGSPGS


■ ■ AMAAMA

■ ■ RPLRPL

■ ■ RIFRIF

■ ■ Severe Male Factor infertilitySevere Male Factor infertility

■ ■ SETSET

■ ■ Patients request Patients request (unknown risk factors)(unknown risk factors)



Since 2004 eleven RCTsSince 2004 eleven RCTs

10 x blastomere10 x blastomere 1 x blastocyst stage1 x blastocyst stage



PGSPGS for for AMAAMA


■■ Current evidence Current evidence do not supportdo not support the use of PGS the use of PGS (aneuploid screening) to increase live birth rates (aneuploid screening) to increase live birth rates in women of advanced material age due to :in women of advanced material age due to :

- increased rate of mosaicism at cleavage stage- increased rate of mosaicism at cleavage stage

- tested cell(s) not necessarily representative of embryo- tested cell(s) not necessarily representative of embryo

- inability to examine all of the chromosomes by FISH….- inability to examine all of the chromosomes by FISH….

■■ Most of the published studies on PGS refer to AMAMost of the published studies on PGS refer to AMA


■■ Even with array CGH limitations regarding ploidy still Even with array CGH limitations regarding ploidy still exist…..exist…..



However, even Array CGH has its own limitations However, even Array CGH has its own limitations regarding cost, experience and ploidy difficultiesregarding cost, experience and ploidy difficulties


Currently there are no conclusive data on the Currently there are no conclusive data on the application of PGS regarding application of PGS regarding - repeated miscarriage,- repeated miscarriage, - implantation failure and- implantation failure and - severe male factor- severe male factor..

In view of the lack of evidence for the effectiveness In view of the lack of evidence for the effectiveness of PGS and the accumulating evidence for its of PGS and the accumulating evidence for its harmfulness harmfulness it is probably unethicalit is probably unethical to perform to perform additional RCTs using cleavage stage biopsy.additional RCTs using cleavage stage biopsy.

________________________________

However in patients with RPL and foetuses with an However in patients with RPL and foetuses with an abnormal chromosomal pattern PGS maybe indicatedabnormal chromosomal pattern PGS maybe indicated




■■ Suggestions for improving resultsSuggestions for improving results

- alternative biopsy timing - alternative biopsy timing (PB and/or trophectoderm)(PB and/or trophectoderm)

- application of technologies for more comprehensive - application of technologies for more comprehensive testing to include all chromosomes testing to include all chromosomes (CGH microarray-based testing)(CGH microarray-based testing)



ESHRE task force onESHRE task force on PGS PGS


■■ In order to clarify if PGS is beneficial, anIn order to clarify if PGS is beneficial, an RCT in AMA patients has been set up using:RCT in AMA patients has been set up using:

- Polar body biopsy- Polar body biopsy

- Array-comparative genomic hydridization- Array-comparative genomic hydridization


________________________________

…………hoping that the collected data will clarify the effectiveness of PGS hoping that the collected data will clarify the effectiveness of PGS

PGDPGD for inherited predisposition to cancer for inherited predisposition to cancer


● ● Familial adenomatous polyposis APCFamilial adenomatous polyposis APC

● ● Li-Fraumeni syndrome p53Li-Fraumeni syndrome p53

● ● Multiple endocrine neoplasia type I MEN1Multiple endocrine neoplasia type I MEN1

● ● Neurofibromatosis type 1 NF1Neurofibromatosis type 1 NF1

● ● Retinoblastoma RB1Retinoblastoma RB1

● ● Von Hippel-Lindau sindrome VHLVon Hippel-Lindau sindrome VHL

● ● Breast & Ovarian cancerBreast & Ovarian cancer

● ● Increasing number of indications!!Increasing number of indications!!


RECOMMENDATIONS:RECOMMENDATIONS: PGDPGD


■■ Before PGD is performed, Before PGD is performed, genetic counseling must be providedgenetic counseling must be provided to ensure that patients fully understand the risk for having an to ensure that patients fully understand the risk for having an affected child, the impact of the disease on an affected child, affected child, the impact of the disease on an affected child, and the limitations of available options that may help to avoid and the limitations of available options that may help to avoid the birth of an affected child.the birth of an affected child.

■■ PGD can reduce the risk for conceiving a child with a genetic PGD can reduce the risk for conceiving a child with a genetic abnormality carried by one or both parents if that abnormality abnormality carried by one or both parents if that abnormality can be identified with tests performed on a single cell.can be identified with tests performed on a single cell.

■■ Prenatal diagnostic testing to confirm the results of PGD is Prenatal diagnostic testing to confirm the results of PGD is encouraged strongly because the methods used for PGD have encouraged strongly because the methods used for PGD have technical limitations that include the possibility for a false technical limitations that include the possibility for a false negative result. negative result.


RECOMMENDATIONS:RECOMMENDATIONS: PGSPGS


■■ Before PGS is performed, Before PGS is performed, thorough education and counselingthorough education and counseling must be provided to ensure that patients fully undersand the must be provided to ensure that patients fully undersand the limitations of the technique, the risk of error and the lack of limitations of the technique, the risk of error and the lack of evidence that PGS improves live-birth rates as currently evidence that PGS improves live-birth rates as currently performed on all the conditions stated earlier performed on all the conditions stated earlier (AMA, RPL, RIF, SET, SMFI etc) except in specific cases.(AMA, RPL, RIF, SET, SMFI etc) except in specific cases.

■■ Decisions concerning future treatment should not be based on Decisions concerning future treatment should not be based on the results of PGS in one or more cycles.the results of PGS in one or more cycles.



CONCLUSION CONCLUSION


Yes in…Yes in…



Single gene disorders Single gene disorders HLA matched sibling embryos to treat genetic conditionsHLA matched sibling embryos to treat genetic conditions of affected childrenof affected children

Carriers of chromosomal abnormalities Carriers of chromosomal abnormalities

RPL when aborted foetuses are chromosomally abnormal !! RPL when aborted foetuses are chromosomally abnormal !!

______________________________At the moment all the other indications need At the moment all the other indications need to be put on holdto be put on hold before we start routinely applying such a costly and before we start routinely applying such a costly and technologically advanced technique with many inherent and technologically advanced technique with many inherent and external limitations external limitations

Future of PGD/PGSFuture of PGD/PGS


BiopsyBiopsy

Polar bodyPolar body Blastocyst biopsyBlastocyst biopsy VitrificationVitrification


PGDPGD

New technology to allow diagnosisNew technology to allow diagnosis Whole genome amplificationWhole genome amplification SNP arrays and array CGHSNP arrays and array CGH

PGSPGS

PCT to see if valid procedure with clinical significancePCT to see if valid procedure with clinical significance NOT cleavage stage biopsyNOT cleavage stage biopsy Try polar body or trophectodermTry polar body or trophectoderm NOT FISHNOT FISH Try arrays (either SNP or array CGH)Try arrays (either SNP or array CGH)

J.HarperJ.HarperAthens PGD Symposium, Athens PGD Symposium, MAY 29-30/2010MAY 29-30/2010

KaryomappingKaryomapping


A Universal method for genome wide analysis of A Universal method for genome wide analysis of genetic disease based on mapping crossovers genetic disease based on mapping crossovers between parental haplotypesbetween parental haplotypes


Handyside AHandyside AAthens PGD Symposium, Athens PGD Symposium, MAY 29-30/2010MAY 29-30/2010

High density genome wide single nucleotide High density genome wide single nucleotide polymorphisms (SNP) genotyping of proband polymorphisms (SNP) genotyping of proband parents and appropriate family member(s) to parents and appropriate family member(s) to establish phase.establish phase.

Mendelian analysis and Karyotyping of the parental Mendelian analysis and Karyotyping of the parental and grandparental haplotypes for each chromosome and grandparental haplotypes for each chromosome or chromosome segment to r-chromosomeor chromosome segment to r-chromosome

ConclusionsConclusions


Karyomapping combines accurate analysis of single gene Karyomapping combines accurate analysis of single gene defects and chromosome aneuploidies and partial deletions defects and chromosome aneuploidies and partial deletions with their parental originwith their parental origin


Handyside AHandyside AAthens PGD Symposium, Athens PGD Symposium, MAY 29-30/2010MAY 29-30/2010

The prevalence of chromosomal abnormalities in patients’ The prevalence of chromosomal abnormalities in patients’ embryos underlines the importance of combined analysisembryos underlines the importance of combined analysis for PGDfor PGD

validation of new technologies !!!validation of new technologies !!!

MARINOS TSIRIGOTISMARINOS TSIRIGOTISMD.FRCOGMD.FRCOGDIRECTOR “AKESO” FERTILITY CENTERDIRECTOR “AKESO” FERTILITY CENTER

THANK U FOR LISTENINGTHANK U FOR LISTENING

Video…blastomere biopsyVideo…blastomere biopsy

Video…polar body biopsyVideo…polar body biopsy

Video…trophectoderm biopsyVideo…trophectoderm biopsy


ReferencesReferences


Preimplantation Genetic Screening:Polar body Screening Study Launched. ScienceDaily (june29.2009)

Preimplantation Genatic Diagnosis May Pose Neurological Risks. Science Daily(july 22,2009) Yu et al; Evaluation of blastomere biopsy using a mouse model indicates the potential high risk of neurodegenerative disorders in the offspring. Molecular & Cellular Proteomics, 2009

Embryo biopsy does not effect early growth and risk of congenital malformations in PGD/PGS babies. ScienceDaily (July 10, 2008).Adapted from materials provided by European Society for human Reproduction & Embryology, via EurekAlert!, a serviceof AAAS.

Embryo biopsy safe for singleton pregnancies, larest study of PGD children suggests. ScienceDaily (Dec. 23, 2009) Adapted from materials provided by European Society for human Reproduction & Embryology, via EurekAlert!, a serviceof AAAS.

Single-cell DNA and FISH analysis for application to preiplantation genetic diagnosis. Chong SS.et al; Currr Protoc Hum Genet. 2010 Jan.

What next for preimplantation genetic screening? More randomized controlled trials needed? Mastenbroek S et al; Hum Reprod. 2008 Dec.

IVF/ICSI with or without preimplantation genetic disorders:a systematic review and meta-analysis. Checa MA et al; J Assist Reprod Genet. 2009 May.

Preimplantation genetic diagnosis and chromosome analysis of blastomeres using comparative genomic hybridization. Wilton L Hum Reprod Update. 2005 Jan.

FISH analysis of 15 chromosomes in human day 4 and 5 preimplantation embryos: the added value of extended aneuploidy detection. Baart EB et al; Prenat Diagn. 2007 Jan.

What next for preimplantation genetic screening? High mitotic chromosome instability rate provides the biological basis for the low succedd rate. Vanneste E et al; Hum reprod. 2009 Nov.

Preimplantation genetic screening for abnormal number of chromosomes (aneuploidies) in in vitro fertilization or intracytoplasmic sperm injection. Twisk M et al; Cochrane Database Syst Rev. 2006 Jan.




What next for preimplantation genetic screening? Harper J et al; Hum Reprod. 2008 Mar.

Is intracytoplasmic sperm injection itself an indicaton to perform preimplantation genetic diagnosis (PGD)? About PGD, invasive prenatal diagnosis and genetic sonography. Ludwing M et al; Fetal Diagn Ther. 2001 Mar-Apr.

Advanced maternal age as an indication for preimplantation genetic diagnosis (PGD)- the need for more judicious application in clinically assisted reproduction. Heng BC. Prenat Diagn. 2006 Nov.

Preimplantation genetic diagnosis. Geraedts JP etal; Clin Genet. 2009 Oct.

Hereditary cancer predisposition syndromes and preiplantation genetic diagnosis:where are we now? Konstantopoulou I et al; JBUON. 2009 Sep.

Attitudes of high-risk women toward preimplantation genetic diagnosis. Quinn G et al; Fertil Steril. 2009 Jun.

Conflict between values and technology: perceptions of preimplantation genetic among women at increase risk for hereditary breast and ovarian cancer. Quinn GP et al; Fam Cancer. 2009 Jun.

Current controversies in prenatal diagnosis 1: Is aneuploidy testing by PGD indicated for all infertile patients undergoing IVF? Harper JC et al; Prenat Diagn. 2009 Dec.

What next for preimplantation genetic screening? A clinician’s perspective. Yakin K, Urman B. Hum Reprod. 2008 Aug.

Preimplantation genetic screening in women of advancsed maternal age caused a decrease in clinical pregnancy rate: a randomized controlled trial. Hardarson T, et al; Hum Repod. 2008 Dec.

RE-analysis of 166 embryos not transferred after PGS with advanced reproductive maternal age as indication. Hanson C et al; Hum Reprod. 2009 Nov.

A prospective randomized controlled trial of preimplantation genetic screening in the “good prognosis” patient. Meyer LR et al; Fertil Steril. 2009 May.




Clinical application of comprehensive chromosomal screening at the blastocyst stage. Schoolcraft WB et al; Fertil Steril. 2009 Nov.

Preclinical validation of a microarray for full modecular karyotyping of blastomeres in a 24-h protocol. Johnson DS et al; Hum Reprod. 2010 Apr.

Use of comprehensive chromosomal screening for embryo assessment: microarrays and CGH. Wells D et al; Mol Hum REprod. 2008 Dec.

Reduction of the multiple pregnancy rate in a preimplantaion genetic diagnosis programme after introduction of single blastocyst transfer and cryopreservation of blastocyst biopsied on day3. El-Toukhy T et al; Hum Reprod. 2009 Oct.

Pregnancy after trophectoderm biopsy of frozen-thawed blastocyst. Lathi RB Behr B. Fertil Steril. 2009 May.

Pregnancies and live births after trophectoderm biopsy preimplantation genetic testing of human blastocyst. McArthur SJ et al; Fertil Steril. 2005 Dec.

Preimplantation genetic diagnosis: state of the art. Basille C e tal; Eur J Obstet Gynecol Reprod Biol. 2009 Jul.Embryo development characteristics in Robertsonian and reciprocal translocations : a comparison of results with non-translocatin caese. Findikli N et al; Reprod Biomed Online 2003 Nov.

Guidelines for good practise in PGD: programme requirements and laboratory quality assurance. Preimplantation Ganetic Diagnosis International Society (PGIS). Reprod Biomed Online. 2008 Jan.

Growth and health outcome of 102-2-year-old children conceived after preimlpantation genetic diagnosis or screening. Desmyttere S et al; Early Hum Dev. 2009 Dec.

Preimplantation genetic diagnosis:state of the art. Basille C et al; Eur J Obtet Gynecol reprod Biol. 2009 Jul.

Preimplantation genetic diagnosis :technology and clinical applications. Swanson A et al;2007 May




ΤΗΕ ΤΗΕ ENDEND

presentation instabul +videos 1 6-2010 (1)

Documents

genetic parents

linked disorders

autosomal dominant disorders

autosomal dominant conditions

gene mutation

female carriers of x

embryos blastomeres

specific mutation