Assisted reproduction.

Fetal
membranes. Fetal develoment.
Twinning. Critical periods of
gestation (5.8)
MF III year

REPRODUCTION AND
INHERITANCE
 Contents
• Fetal membranes.
• Artificial fertilization and its methods.
• Multiple pregnancy. Monozygotic and
dizygotic twins. Siamese twins
• Fetal developmental period.
Organogenesis. Critical periods of
gestation. Etiology and classification of
developmental defects.
 Fertilization and
Assisted reproduction
techniques
 Fertilization
A chain of events:
• Sperm capacitation
• Sperm – zona pellucida binding
• Acrosome reaction
• Penetration of the zona pellucida
• Sperm-oocyte binding
• Egg activation and cortical reaction
• Zona reaction
• Post-fertilization events
 Acrosome reaction
• Binding to ZP3, activates G protein on the
surface of the sperm, opens the Ca++ channels
and causes the fusion of plasma membrane of
the sperm with the outer acrosomal membrane.

Developmental Biology, 8th ed.

• Insemination is a necessary prerequisit
condition for fertilization to occur.
• Insemination in humans is associated with
interpersonal relationships which lead to
sexual intercourse, during which the male
gamets are delivered into female vagina.
 Methods of
assisted reproduction
Some form of external fertilization may be
necessary when a man and woman wish to
have a child but one or the other is
biologically incapable of contributing to the
normal process of internal fertilization.

• In vivo fertilization - IUI

• In vitro fertilization, including ICSI.
Intra uterine insemination (IUI)
 In vitro fertilization (IVF)
In 1978, the first human birth following in vitro fertilization occurred in
England

1) Stimulation of ovaries for increased oocyte growth and

maturation, but inhibiting natural ovulation

2) Harvesting of oocytes

3) Insemination with processed

sperms in a Petri dish
or intracytoplasmic sperm
injection (ICSI),
if sperm numbers are very small
Intracytoplasmic sperm injection (ICSI)
 In vitro fertilization (IVF)
4) Growth of embryos in an incubator
Allowing the embryos to grow in an environment established by
culturing other cells from the woman sometimes improves the quality of
the embryos (co-culture).
5) Once the embryos mature, a number (1-3) of them are
inserted through the cervix into the uterus by means of a
small catheter. Some or all of the embryos may undergo assisted
hatching to increase implantation rates.
6) Supernumerous embryos are cryopreserved.
7) Harvested oocytes may be cryopreserved before or
after fertilization in pronucleus stage.
www.youtube.com/watch?v=V6-v4eF9dyA
Cryopreserved Embryos
 In vitro fertilization (IVF)
• Opportunity to alleviate infertility from a
variety of causes
• High costs - low rate of success;
• Multiple births
• Potential* to karyotype fertilized ova and
help to prevent chromosomal
abnormalities, sex-linked disorders;

*Not accepted in multiple countries over the world.

Preimplantation diagnostics
 Embryogenesis

From fertilization to the end of

8th week (56 days); it equals to the
10th week of pregnancy
 Periods of embryogenesis

Zygote – the first mitotic division after 24-30 hrs. after fertilization
Segmentation – morula formation
Blastogenesis – inner cell mass and trophoblast
Gastrulation – three germinal layers –
ecto-, meso- and endoderm
Histogenesis – tissues differentiate
Neurulation – formation of neural tube
Organogenesis – budding of most organs
 The Carnegie Stages of Early Human
Development
• Wilhelm His – pioneer of reconstructive 3D
views of embryos – the end of 19th century.
• Franklin P. Mall, established Embryo
collection at Carnegie Institute, Baltimore,
he was the first to distinguish human
developmental stages (1914).
• Ronan O'Rahilly continued formulation of
Carnegie stages and announced detailed
description in 1972.
Length
mm days
Stage lengh mm days
 Embryogenesis continues into
fetogenesis at day 56

• The end of differentiation of gonads.

• The embryo is 39-40 mm long and has
appearance of human
• All organ primordia have already formed
• External genitals are undifferentiated
 Fetogenesis
from day 57 until delivery
 During fetogenesis:
• Growth in length and weight gain;
• Changing proportions of head, body
and extremities;
• more distinct features of the face;
• Future growth and functional
development of organs and tissues,
Stronger relation of nerves and
muscles, active movements, reflexes
The 4th month
• External genitals
are distinctive;
• More active
movements, but
rarely felt by the
mother before the
week 17,
• Length of the
fetus reaches
16 cm
 20 weeks
• Fetus length 25-26
cm,
• weight 280-300 g.

• Red blood cells are

mainly produced in
the bone marrow.
• Breathing is not
possible at this time
 23-26 weeks
• Length 30 cm, weight 500-800 g.

• If born at this time, the newborn has 50%

chance of survival.
 28 weeks
• Length 35 cm, weight 1000-1200 g.
• Reacts to light and sound.
• If born at this time, the newborn can
breathe, cries faintly, moves actively his
extremities.
 32 weeks

• Length 40-42 cm, weight 1500-1700 g.

• Testes descend into scrotum.
 36 weeks
• The fetus, when born, cries aloud, has
sucction reflex.

40 weeks
• The fetus is 49-52 cm long and weighs
more than 2500 g
• The skin is smooth, pink, with tiny hair on
the shoulder region. His head is 1/4th of
the body length. Nails are longer than the
fingers. Such fetus is completely mature.
Site of
production Placenta
Hormone concentration

Estrogen:
Estriol
in mother‘s blood

Estrone
Estrogen (estradiole)

estradiole

Weeks
 After delivery
• After separation from the mother the fetus
is called a newborn (Neonatus).
• When the placental circulation is
compromised, the neonate inspires for the
first time.
• Cardiovascular and pulmonary system
undergo dramatical changes after birth.
 After delivery
• With inspiration, the circulation in the
pulmonary vessels starts. Foramen ovale
closes due to increase presure in the left
atrium.
– Truncus arteriosus is gradually
obliterated.
– Umbilical arteries and the vein are
obliterated and become corresponding
ligaments
– Heart rate - 110 bpm.
Determination of age of
embryo and fetus
Prenatal diagnostics
 I. According to length of the embryo

C-R (Crown-rump); C-H (Crown-heel); A-R - (atlas-rump)

Length
In milimeters

Weeks of gestation
II. According to the time
of appearance of somites
Days Number Days Number
of somites of somites
17 1-3 25 24-26
18 3-5 26 27-28
19 6-8 27 29-30
20 9-11 28 30-31
21 12-14 29 32-33
22 15-17 30 34-35
23 18-20 31 36-37
24 21-23 32 38-39
 The age of embryo also may be
determined by Carnegie stages,
each of which have characteristic
morphologic criteria.
Haase’s equation for determination of gestational
age according to fetal length
Month of Assessment Embryo/fetus
gestation length in cm
I 1x1 1
II 2x2 4
III 3x3 9
IV 4x4 16
V 5x5 25
VI 6x5 30
VII 7x5 35
VIII 8x5 40
IX 9x5 45
X 10x5 50
 Age of gestation
The duration of clinical gestation is
calculated from the first day of the last
menstrual cycle until delivery;

In embryology the gestational age is

calculated from the moment of
ovulation and fertilization
Maturation of the fetus according to
gestational age
• Duration of normal pregnancy is 280 days, or 40
weeks, gestational age = 280-14=266 days.
• Full-term neonate is delivered in the week 38 to
42 from the onset of the last menstrual period,
pre-term baby is born before the end of the 37th
week of gestation; postmature baby is born
after the week 42.
• Mature baby is considered, when the length is
>49 cm, body weight exceeds 2,5 kg, and there
are other signs of morphological maturity.
Signs of morphological maturity
• Length (>49 cm)
• Weight (>2500g; usually 3000-3500g)
• Head circumference 35 cm;
• Position of external genitals;
• Skin thickness and the length of hair;
• position of ears, their cartilage flexibility.
• Length and hardness of nails;
• Vernix caseosa; lanugo hairs;
• Shape of chest and abdomen
 Estimation of the age of the fetus
• CRL at the age of 7-14 weeks may be
determined by ultrasound
• At the age over 16 weeks biparietal
diameter, head and abdominal
circumferences, femur length are
measured
Correlation of embryonic and fetal age, weight and length

Gestational age
Length (CRL) cm Weight , g
weeks
 Functional maturity
Signs Score 0 Score 1 Score 2

Heart rate No Less than More than

100bpm 100bpm
breathing No Rare, irregular Regular with
cry
Muscle Weak Extremities Active motions
tone slightly bend
reflexes No Grimace Coughing,
sneezing
Skin Pale to blue Trunk - pink, The whole
colour extremities - blue body is pink
 Prenatal diagnostics (fetology)
• Maternal serum screening for:
– Alpha-fetoprotein; hCG; uE3; Inhibin-A
• Ultrasonography
• Amniocentesis
• Chorionic villus sampling
– (alternatively fetus DNA analysis may be
performed on syncytiotrophoblast nuclei,
obtained from mother‘s blood sample)
Twinning, or multiple pregnancy
• Monozygotic (homozygotic) twins develop
from one egg-cell, fertilized with one
sperm cell; ¼ - 1/3.
• Dizygotic (heterozygotic) twins (duplets,
triplets, etc.) - develop from separate
eggs, fertilized with different sperms; 2/3 –
3/4. There may be two follicles ovulating or
one follicle releasing two oocytes.
 Twinning is an example of
blastopathies
• When monozygotic twins develop from
one egg-cell, fertilized with one sperm cell.
Twinning from one zygote
Conjoined twins

Result when splitting occurs in late blastocyst,

in a bilaminar germinal disk,
or there is formation of multiple primitive streaks.
Teratology (Greek teratos – monster),
is a part of embryology, dealing with malformations,
their causes and mechanisms of development.

Endogenous factors: Exogenous factors:

genetic or biological, chemical,
chromosomal physical,
anomalies Interaction of environmental
endogenous
and exogenous
factors
Congenital and heritable Non-heritable
 Critical periods of gestation
• I – from conception until implantation –
blastogenesis
• II – 3 - 8 weeks of gestation –
embryogenesis
• III – fetogenesis until birth
 I
• The time from conception until implantation
known as the "all or none" period,

• preimplantation period, blastogenesis – until 16

days after conception. Blastula is very sensitive
– blastopathies:
– Ectopic implantation,
– Twins (conjoined);
– 1/3 of fertilized ova perish unnoticed.
 II
• The embryonic period, from 17 to 56 days after
conception is the period when the basic steps in
organogenesis occur. This is the period of maximum
sensitivity to teratogenicity since not only are
tissues differentiating rapidly but damage to them
becomes irreparable. Exposure to teratogenic agents
during this period has the greatest likelihood of causing a
structural anomaly. Since teratogens are capable of
affecting many organ systems, the pattern of anomalies
produced depends upon which systems are
differentiating at the time of teratogenic exposure. The
defects are gross and are called embryopathies (e.g.
agenesis of an organ).
 III
• The fetal phase, from the end of the embryonic stage to
term, is the period when growth and functional
maturation of organs and systems already formed
occurs. Teratogen exposure in this period will affect fetal
growth (e.g., intrauterine growth retardation), the size of
a specific organ, or the function of the organ, rather
than cause gross structural anomalies. The term fetal
toxicity - fetopathy. Of particular interest is the
potential effect of psychoactive agents (e.g.,
antidepressants, antiepileptics, alcohol and other drugs
of abuse) on the developing central nervous system,
which has led to a new field - behavioural
teratology.
• Many organ systems continue structural
and functional maturation long after birth.
Most of the adenocarcinomas associated
with first trimester exposure to the
synthetic estrogen, diethylstilbestrol,
occurred many years after the exposure.
Fetal membranes
• Human fetal membranes, membranae
fetales humana, are all the structures
which participate in the fetal development
and are expelled from the uterus during
delivery of the fetus
- The yolk sack,
- chorion,
-placenta,
- amnion,
- alantois,
- umbilical cord.
 Placenta –
the place where chorion villosum (pars fetalis)
meets the decidua basalis (pars materna)

Its functions:
- gas exchange,
- exchange of nutrients and waste products,
- immunologic defense ,
- accumulation of certain vitamins (A, D etc.), iron
and glycogen,
- synthesis of hormones, enzymes
Cytotrophoblast (CT) cells Inactivation Activation
through through
form the multinucleated conjugation in deconjugation in
external layer of placental villi fetal adrenal placenta
– the sincytiotrophoblast, cortex and liver
Estriol,
which is supplemented with estrone,
estradiol
new CT cells. secreted
Sincytiotrophoblast is into
maternal
important for gas and blood
substances exchange, as well
as for endocrine activity:
chorionic gonadotropin (CG)
and somatomammotropin
(placental lactogen (PL); Progenitor
progesterone and estriol substances
from mother
(cholesterol,
pregnenolone)
Cytotrophoblast cells
of the stem villi
penetrate the uterine
mucosa, migrate to
spiral arteries and
overgrow their inner
and muscular layer to
ensure continuous
flow of maternal blood
into intervillous
spaces
 Exchange of substances through the placental barrier
Substances Examples
Beneficial
Gases Oxygen, carbon dioxide
Nutrients Glucose, amino acids, free fatty acids, vitamins
Metabolites CO2, urea, uric acid, bilirubin, creatine, creatinine
Electrolytes Na+, K+, Cl-, Ca2+, PO42-
Erythrocytes Fetal and maternal both (a few)
Maternal serum proteins Serum albumin, some protein hormones (thyroxin, insulin)
Steroid hormones Cortisol, estrogen (unconjugated only)
Immunoglobins IgG (confers fetal passive immunity)
Harmful
Poisonous gases Carbon monoxide
Infectious agents Viruses (HIV, cytomegalovirus, rubella, Coxsackie, variola,
varicella, measles, poliomyelitis), bacteria (tuberculosis,
Treponema), and protozoa (Toxoplasma)
Drugs Cocaine, alcohol, caffeine, nicotine, warfarin,
trimethadione, phenytoin, tetracycline, cancer
chemotherapeutic agents, anesthetics, sedatives,
analgesics
Immunoglobins Anti-Rh antibodies
 Exchange of substances
• Active: • Passive:
– Amino acids; – gases
– Salts – water,
– Glucose – bilirubin (unconjugated
– Vitamins passes the placental
barrier very well)

Transcytosis:
• proteins (IgG)
During implantation decidual reaction
is observed – fibroblasts in the uterine
stroma differentiate into large polygonal
cells – decidual cells.
They secrete: decidual prolactin (dPRL),
insulin-like growth factor binding protein-1
(IGFBP-1).
Decidual cells in the pars materna
 Decidual cell functions
- trophic (at the beinning of implantation they
possess nutrients, glycogen, lipids),
- secretion (secrete various citokines, which
have influence on the attachment of trophoblast
in the uterine mucosa),
- protective (secrete MMP inhibitors, interstitial
ground substance and adhesion molecules,
which prevent the sincytiotrophoblast from
invasion too deep into endometrium).
 Placental barrier

Initially

Before delivery
 Mature placenta
(before delivery)
• Discoid shape
• 15-20 cm in diameter
• 500 g weight
• 15-20 cotyledons
• Umbilical cord is attached near the center,
with bloods vessels gradually penetrating
the chorionic plate, branching and entering
into the villi
 The amniotic cavity is filled with clear
watery fluid, amniotic fluid, produced by
amniotic cells
(amount increases from 30 ml /10 weeks to
450ml /20 weeks and
800-1000ml /37 weeks.)
Thanks for your attention!