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The Chick Embryo Amnion As an in Vitro Culture System for Ivf and Nt Embryos

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The Chick Embryo Amnion As an in Vitro Culture System for Ivf and Nt Embryos Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2004 The chick mbre yo amnion as an in vitro culture system for IVF and NT embryos Tonya Renea Davidson Louisiana State University and Agricultural and Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Animal Sciences Commons Recommended Citation Davidson, Tonya Renea, "The chick mbre yo amnion as an in vitro culture system for IVF and NT embryos" (2004). LSU Doctoral Dissertations. 1280. https://digitalcommons.lsu.edu/gradschool_dissertations/1280 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. THE CHICK EMBRYO AMNION AS AN IN VITRO CULTURE SYSTEM FOR IVF AND NT EMBRYOS A Dissertation Submitted to the Graduate Faculty of Louisiana State University and Agricultural and Mechanical College In partial fulfillment of the Requirements for the degree of Doctor of Philosophy in The Interdepartmental Program in Animal and Dairy Sciences by Tonya Renea Davidson B.S., Oklahoma State University, 1997 M.S., Oklahoma State University, 2000 December 2004 ACKNOWLEDGEMENTS I would like to begin by dedicating this dissertation to my mom, Linda Millspaugh. Mom, you have always been my role model, my support system and my foundation. You taught me that I could be whatever I wanted to be, and you had faith in me even when I didn’t have faith in myself. I am so proud to be your daughter. I would also like to dedicate this dissertation to my brother, Casey Millspaugh. Thank you Casey, for always believing in me and looking up to me. That has been more encouragement than you will ever know. I would like to express me sincere gratitude to my major professor, Dr. Robert Godke. Thank you for allowing me to pursue my Ph.D. at LSU. You have provided me with a firm scientific background, but more importantly, you have encouraged me to stand up for that which I believe. You have been a wonderful mentor and I appreciate all that you have done for me. To Dr. Leibo, Dr. Lynn and Dr. Williams. Thank you so much for serving on my committee, and for all of the assistance you have provided to make my time at LSU such a success. It has been a great pleasure working with all of you. I have learned so much from each of you and I am a better scientist for having known you. To Trey Chapman, Kyle Hebert, Laurie Henderson, Andrea Huval, Angela “Beanie” Klumpp, Luke Lenard, Alison Mosian, Kyle Hebert, Dr. Masao Murakami and James “Par” Sterling. Thank you for all that you have done for me. You have each touched my life in a very special way and I will always remember you. To Kristy Bruce. Thank you for saying “Hello” that first day of class. Since that time, you have been a wonderful source of entertainment and support, but more importantly, you have been an exceptional friend. I am so much better for having known you. And finally, to Edward Ferguson. My words can not adequately express how I truly feel for you. You are my best friend and the love of my life. You have never left my side, no matter how hard things got, and for that I will always love you. ii TABLE OF CONTENTS ACKNOWLEDGEMENTS ................................................................................................. ii LIST OF TABLES..............................................................................................................vi LIST OF FIGURES..........................................................................................................viii ABBREVIATIONS USED IN THE DISSERTATION..........................................................xi ABSTRACT......................................................................................................................xii CHAPTER I. INTRODUCTION .................................................................................... 1 CHAPTER II. LITERATURE REVIEW .......................................................................... 3 In Vitro Production of Embryos.............................................................................. 3 Production of Bovine Embryos by In Vitro Fertilization........................... 3 Production of Bovine Embryos by Nuclear Transfer............................... 4 In Vitro Embryo Culture .......................................................................... 6 Influence of In Vitro Production on Embryo Development................................... 11 Embryo Development Rates................................................................. 11 Embryo Morphology ............................................................................. 12 Pregnancy and Calving Rates .............................................................. 14 Large Offspring Syndrome and Associated Disorders.......................... 16 Apoptosis ............................................................................................. 18 Chick Embryo Co-Culture.................................................................................... 24 Biochemistry of the Avian Egg.............................................................. 24 Early Use of Chick Embryo Extracts in Mammalian Cell Culture ......... 25 In Vitro Culture of Chick Embryos ........................................................ 26 Chick Embryo Co-Culture..................................................................... 26 CHAPTER III. CULTURING IN VITRO PRODUCED BOVINE EMBRYOS IN THE AMNIOTIC CAVITY OF THE DEVELOPING CHICKEN EMBYRO...... 28 Introduction.......................................................................................................... 28 Materials and Methods........................................................................................ 29 Experimental Design ............................................................................ 29 Experimental Methods.......................................................................... 31 Statistical Analysis................................................................................ 36 Results ................................................................................................................ 36 Discussion........................................................................................................... 41 CHAPTER IV. USE OF THE AMNIOTIC CAVITY OF THE DEVELOPING CHICK EMBRYO FOR CULTURING BOVINE 8-CELL EMBRYOS DERIVED FROM SOMATIC CELL NUCLEAR TRANSFER ................................. 46 Introduction ......................................................................................................... 46 Materials and Methods........................................................................................ 47 Experimental Design ............................................................................ 47 Experimental Methods.......................................................................... 49 iii Statistical Analysis................................................................................ 53 Results ................................................................................................................ 53 Discussion........................................................................................................... 54 CHAPTER V. THE EFFECT OF ALTERATIONS IN THE NORMAL ANGIOGENIC PATTERN OF THE CHICK CHORIOALLANTOIC MEMBRANE ON BOVINE EMBRYO DEVELOPMENT AFTER CHICK EMBRYO CO-CULTURE...................................................................................... 60 Introduction ..................................................................................................... 60 Materials and Methods .................................................................................... 61 Experimental Design ............................................................................ 61 Experimental Methods.......................................................................... 63 Statistical Analysis................................................................................ 69 Results............................................................................................................ 69 Discussion....................................................................................................... 78 CHAPTER VI. DEVELOPMENT OF 8-CELL BOVINE EMBRYOS IN CULTURE MEDIUM SUPPLEMENTED WITH CHICK AMNIOTIC FLUID (CAF).. 80 Introduction...................................................................................................... 80 Materials and Methods.................................................................................... 81 Experimental Design ............................................................................ 81 Experimental Methods.......................................................................... 81 Statistical Analysis................................................................................ 85 Results ............................................................................................................ 86 Discussion....................................................................................................... 86 CHAPTER VII. ESTABLISHMENT AND CULTURE OF CELLS DERIVED FROM DAY-6 CHICK EMBRYOS.................................................................... 93 Introduction.........................................................................................................
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