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ÊéÃûAtlas of
PREIMPLANTATION
GENETIC DIAGNOSIS
×÷ÕßAnver Kuliev, MD, PhD
Svetlana Rechitsky, PhD
and
Oleg Verlinsky, MS
Reproductive Genetics Institute
Chicago, Illinois, USA
Ò³Êý324
³ö°æÉç CRCpress
International Standard Book Number-13: 978-1-4665-9840-9 (eBook - PDF)
Acknowledgments ii
Preface v
Introduction vi
SECTION Ia•¡­ Review of Current Methods and Experience in Preimplantation
Genetic Diagnosis
1 Normal and abnormal human preimplantation development in relation to preimplantation genetic
diagnosis and establishment of embryonic stem cell lines 1
Introduction 1
Oocytes at different stages of meiosis I 1
Oocytes at metaphase II and variations in first polar body formation 2
First polar body morphology in relation to chromosomal abnormality in oocytes 2
Oocytes following fertilization and second polar body extrusion 3
Normally and abnormally cleaving embryos 4
Normal and abnormal blastocyst formation 4
Establishment of human embryonic stem cell lines 5
References 5
2 Micromanipulation and biopsy of polar bodies, blastomeres, and blastocysts 7
First polar body removal 9
Intracytoplasmic sperm injection 10
First and second polar body removal 11
Blastomere biopsy 11
Blastocyst biopsy 12
Laser-assisted hatching 14
Trophectoderm biopsy 14
References 15
3 Nuclear transfer techniques for preimplantation diagnosis and prospect for artificial gamete formation 16
Visualization of polar body and blastomere chromosomes 16
Sperm duplication 18
Development of artificial human gametes in vitro 19
References 20
4 Preimplantation diagnosis for aneuploidies 21
Introduction 21
Preparation of polar bodies, blastomeres, and trophectoderm samples for fluorescence in situ
hybridization analysis (FISH) 22
Preparation of PB1 and PB2 22
Preparation of blastomeres 25
Preparation of biopsied trophectoderm cells 25
Pretreatment, probe application, hybridization, and washing 26
Hybridization procedure 26
Washing procedures 27
Fluorescent signal evaluation 28
Array-based 24-chromosome aneuploidy testing 29
Chromosomal abnormalities in polar bodies 29
Chromosomal abnormalities in cleaving embryos and blastocysts 30
References 31
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