LABORATORY

PRODUCTION OF
ANIMAL
EMBRYOS
BY
ESAM EL-DIN THARWAT, PhD
PROFESSOR OF ANIMAL PHYSIOLOGY,
FACULTY OF AGRICULTURE,
AIN SHAMS UNIVERSITY.
 What are Reasons for In
Vitro Production of Farm
Animals Embryo?
 Reasons for In Vitro
Production of Farm Animals
Embryo
◼ First, this technique provides large numbers of
embryos for commercial transfer and calf
production.
◼ Second, the economic feasibility of embryo
cloning by nuclear transfer requires that the
enucleated oocytes be produced in-vitro from
abattoir recovered ovaries and that the new
zygote be developed in-vitro to a stage suitable
for re-cloning.
 Laboratory Production of
Animal Embryos
◼ Obtain by four methods:
◼ In vitro fertilization.
◼ Imprinting.
◼ Cloning.
◼ Gene transfer.
 1-In vitro Fertilization
◼ In vitro fertilization is the creation of
embryos by mixing sperms and ova in the
laboratory.
◼ A- Female Gametes
Oogenesis ( or development of the
ovum): Takes place in the ovary. This
process is more or less similar to that
of spermatogenesis and includes the
usual three successive phases.
A- Multiplication phase.
B- Growth phase.
C- Maturation phase
 Ovary
Follicle
Corpus luteum
Ruptured follicle
(ovulation) Tertiary follicle Primary follicle

Egg nest

Tertiary
follicle

Atretic follicle Secondary

follicle

Corpus albicans
Corpus luteum
 Follicle Cells
Flattened cells surround oocyte Nucleus

Increase in size of oocyte and

Height of follicular cells Ooplasma
Follicular
Cells

Surrounded by several
layers of follicular cells
Oocyte

Follicle forms a fluid filled cavity

Differentiation of several distinct
cell layers in the follicle wall
Fluid Antrum
Cavity
 Fetal Oocyte Development
Mitosis Meiosis
Fertilization Birth

Cow
280 d

Sow
114 d

Mouse
19 d
 Oogenesis
Primordial Germ Cells

Oogonia
Initiation of Meiosis
Interphase Primary Prophase of Meiosis
Oocyte leptotene
DNA synthesis zygotene
pachytene
Meiotic prophase begins diplotene
dictyotene
Growth Meiotic Arrest
Gonadotropin
Independent zona pellucida formation
mRNA production
protein synthesis
cortical granule formation

Gonadotropin
Dependent nuclear maturation
cytoplasmic maturation
 First polar body
Resumption of emitted, arrest at
LH Surge Meiosis metaphase II of meiosis

4N 4N 2N

Primary Oocyte Sperm

Ovulation Penetration Secondary
Oocyte
• Dog • Dog Ovulation

• Fox • Fox (most species)

Zygote (pronucleate egg)

N N
Sperm
2N Penetration
 Maturation Promoting Factor (MPF)
LH Surge Metaphase Promoting Factor

Primary Oocyte GVBD Metaphase I

MPF MPF

GV-Intact

PB-1

Ovulation
MPF MPF

Metaphase II Secondary Oocyte

Hormonal control of the ovarian function