1

Cloning for Beginners

2

Dolly's lamb

2-1

Megan and Morag

2-2

Cloned sheep

2-3

Cloned cattle

2-4

Cloned sheep

2-5

Cloned pigs

3

Basic biology-Differentiation

3-1

Basic biology-DNA

3-2

Basic biology-chromosomes

3-3

Cattle embryos in different stages of development

3-4

Basic biology-fetus

4

Understanding of differentiation in the 1800s

5

Roux experiment (1850-1924)

6

Driesch experiment (1867-1941)

7

Early conclusions- genetic determinants

8

Primitive cloning

9

Spemenn's ideas on "fantastical experiments"

10

What is nuclear transfer?

11

Nuclear transfer in frogs

12

Nuclear transfer in mammals

13

Nuclear transfer schematic

14-1

MII oocyte

14-2

Enucleation

14-3

Chromosomes

14-4

Oocyte couplet

15

Factors Affecting Nuclear Transfer Embryos

16

Cell cycle interactions in NT embryos

17

Methods of Nuclear Transfer

18

Donor Cell Cycle Stage

19

Early Developmental Events

20

Relationship Between MZT and Development of NT Embryos

21

Explanations of MZT relationship to NT Embryos

22

Nuclear Transfer in Mammals

23

Nuclear Transfer from Somatic Cells

24

NT Objectives

24-1

Day-9 Sheep Blastocyst

24-2

Cell culture from sheep blastocyst

25

NT Cultured ED Cells Oct 1993-Feb 1994

26

NT Cultured ED Cells Oct-Dec 1994

27

Schematic of Cell Cycle

28

Methods of Nuclear Transfer with GO/G1 Cells

29

Nuclear Transfer using Serum Starved ED Derived Cells

30

Nuclear Transfer from Somatic Cells

31

Early Developmental Events

31-1

More-Early Developmental Events

32

Isolation of an Embryo Derived Cell Line (SEC 1)

33

Four male sheep

34

Isolation of Primary Foetal Fibroblasts (BLWF1)

35

Black Welsh Rams

36

Isolation of Adult Mammary Epithelial Cell Line (OME)

37

Cover of Nature

37-1

Photo of Dolly the sheep

39

Donor Cell Cycle Stage

40

Why May Quiescence be Important?

41

Donor Cell Cycle Stage-Senescence

42-1

Other Species-cattle

42-2

Other Species-mice

42-3

Other Species-goats

42-4

Other Species-pigs

43

Losses and Abnormalities

43-1

Losses and Abnormalities-Cartoon

43-2

Losses and Abnormalities-Headlines

43-3

Losses and Abnormalities-Headlines

44

Abnormalities

45

Losses-chart

46

Precise genetic modification

47

Objectives of genetic modification

48

Microinjection technology

49

Limitations of microinjection technology

50

Nuclear Transplantation

51

All Animals Transgenic

52

Instant Production Flocks/Herds

53

Pre-selection for High Expression

54

Gene Removal/Modification

55

Potential Benefits of Nuclear Transfer

56

Major Applications

57

List of Current Projects

58

Xenotransplantation

59

Other Applications

60

Genetic Preservation

62

Requirements Properties & Characteristics Required of Donor Cells

63

Nuclear Transfer Beyond Polly

64

Polly, Holly, Molly & Olly

65

Comparison of pronuclear injection versus Nuclear Transfer

66

NT Lambs Transgenic for ECSOM

67

NT Lamb Transgenic for Calcitonin

69

Gene Targeting in Mouse Cells

70

Objectives in Sheep

71

World's First Targeted Cloned Sheep

72

Gene Targeting Candidates

73

Time Magazine Cover Story-"Clones"

74

Is Cloning Possible in Humans?

75

Cloning Humans-Considerations

75-1

"Boys from Brazil"

76

Cloning in Humans

77

Potential Hazards of Cloning in Humans

78

Uses of Cloning in Humans

79

Sexual Reproduction--A unique Individual

80

Cloning Related Techniques in Human Reproduction

81

Cytoplasmic Inheritance in Humans

83

What is a Stem Cell?

84

In vitro & In vivo differentiation of ES cells

85

Sources of Stem Cells

86

Nuclear Transfer in Stem Cell

87

Human Nuclear Transfer

88

Cloning Technology and Humans (a)

89

Cloning Technology and Humans (b)

90

Production of Chimeras

91

Somatic Cell Nuclear Transfer

92

Problems with Cell Therapies

93

Advantages of autologous Stem Cells via NT

95

"Dedifferentiation" of Cells