Chapter 8

Further development of Embryo

(Embryonic Period:
4–8 Weeks of Development)

Textbook of
HUMAN EMBRYOLOGY
With Clinical Cases and 3D Illustrations
Author: Dr Yogesh Sontakke, JIPMER, Pondicherry

©CBS Publishers & Distributors Pvt Ltd, New Delhi

As per:
Competency based Undergraduate curriculum 1
• AN79.4 Describe the development of somites and intra-embryonic coelom
• AN79.5 Explain embryological basis of congenital malformations, nucleus pulposus, sacrococcygeal
teratomas, neural tube defects
• AN79.6 Describe the diagnosis of pregnancy in first trimester and role of teratogens, alpha-
fetoprotein
Continued in Chapter 8……….

Medical Council of India, Competency based Undergraduate curriculum for the Indian Medical Graduate, 2018. Vol. 1; pg 1-80.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 2

Iductionntro
∙ Embryonic period / organogenic period:
∙ Period of 4–8 weeks of the development.
∙ All derms are changing and assign the fate for further development.
∙ Germ disc get folded to form identifiable shape of human embryo.

∙ Events of embryonic period are grouped as follows:

- Ectodermal differentiation
- Mesodermal differentiation
- Embryonic folding
- Endodermal differentiation

∙ Teratogens (factor causing embryonic malformation) affects the developing embryo to

produce congenital anomalies.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 3

Differentiation of
Ectoderm
∙ At the beginning of the third week, central part of ectoderm overlaying the
developing notochord thickens to form neural plate or neuroectoderm.
∙ Remaining part of the ectoderm is called surface ectoderm.
Neuroectoderm

∙ Neural plate grows to form peripheral neural folds and central depressed
area called neural groove.
∙ Neural folds start fusion in central (cervical) region. Fusion extends
craniocaudally to form neural tube.
∙ Cranial open end of neural tube called anterior neuropore closes by the
25th day.
∙ Caudal open end of neural tube called posterior neuropore closes by the
27th day.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 4

Differentiation of Ectoderm
Neuroectoderm

∙ Cells at the margin of neural plate form the neural crest.

∙ Neural tube gets separated from surface ectoderm, whereas neural crest cells come to
lie between ectoderm and neural tube.
∙ Cephalic part of neural tube shows dilations as forebrain vesicle, midbrain vesicle,
and hindbrain vesicle, whereas distal part of neural tube forms spinal cord.
∙ Neuroectoderm form entire central and peripheral nervous system and autonomic
ganglia.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 5

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 6
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 7
Surface ectoderm
∙ Surface ectoderm shows depression at buccopharyngeal membrane and cloacal
membrane. At these two depressions ectoderm is firmly adherent to the endoderm.
Buccopharyngeal membrane (oropharyngeal membrane):
∙ In the third week, it lies cranial to prechordal plate. On development of head fold, it
comse to lie between forebrain vesicle and pericardial bulging.
∙ At the beginning of the fourth week, buccopharyngeal membrane ruptures to form
communication between the amniotic cavity and primitive gut. (MCQ)
∙ Cloacal membrane divides into anterior urogenital membrane and posterior anal
membrane

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 8

Derivatives of surface ectoderm
Surface ectoderm forms the following structures:
1.Epidermis, hair and nail
2.Sebaceous glands and sweat glands
3.Olfactory pit
4.Optic vesicle and lens vesicle
5.Otic vesicle
6.Branchial clefts
7.Rathke’s pouch
8.Epithelial lining of cheek, gum, teeth enamel, root of mouth, nasal cavity
and paranasal air sinuses
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 9
9.Salivary glands
Mesodermal differentiation

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 10

Mesodermal differentiation
∙ By the end of third week, primitive streak start
regressing.
∙ The mesodermal cells that are migrated from
primitive streak start forming rod-shaped on either
side of the notochord.
∙ Condensed mesodermal rods are separated by
shallow longitudinal grooves into three parts:
A. Paraxial mesoderm: lies on either side of
notochord.
B. Intermediate mesoderm: lies lateral to the
paraxial mesoderm.
C. Lateral plate mesoderm: It is the lateral most
part of the mesoderm.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 11

Mesodermal differentiation
Paraxial mesoderm
∙ Situated on either side of the notochord.
∙ Extent: from the prechordal plate to the
primitive streak.
∙ Form a series of rounded, whorl-like mass
called somitomeres.
∙ The somitomeres further form discrete
block of segmental mesoderm called
somites or metameres.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 12

Mesodermal differentiation
Intermediate mesoderm

∙ Lies between paraxial mesoderm and lateral plate

mesoderm.
∙ It forms kidneys and sex glands (ovaries and testis).
Lateral plate mesoderm

∙ Extent: from the intermediate mesoderm to

extraembryonic mesoderm.
∙ Cranialy it is continued with pericardial bar. (Pericardial
bar is mesodermal condensation cranial to the
buccopharyngeal membrane).
∙ Small cavities appear in lateral plate mesoderm that fuse
to form intra-embryonic coelom.
∙ Similar cavity develops in pericardial bar called
pericardial sac.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 13

Formation of layers of lateral plate mesoderm
∙ Intra-embryonic coelom divides lateral plate
mesoderm into two layers:
A. Somatopleuric layer: parietal layer, lies in
contact with ectoderm.
B. Splanchnopleuric layer: visceral layer, lies in
contact with endoderm.
∙ Continuously growing intra-embryonic coelom
separates somatopleuric and splanchnopleuric
layers laterally and develop communication with
extra-embryonic coelom.
Formation of coelomic cavity
∙ Septum transversum:
Pericardial sac communicates with
∙ It is a cranial mesodermal horseshoe-shaped bar
connecting somatopleuric layer with splanchnopleuric intraembryonic coelom to form
layers. inverted U-shaped tubular passage.
∙ Later forms diaphragm. Coelomic cavity forms pericardial,
pleural and peritoneal cavities.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 14

Fate of lateral plate mesoderm
A. Somatopleuric layer:
- Parietal layers of peritoneal, pericardial and
pleural cavities
- Dermis
- Pectoral and pelvic girdles
- Skeletal elements of limbs
B. Splanchnopleuric layer:
- Visceral layer of pericardial, peritoneal and
pleural cavities
- Musculature and connective tissue of gut,
respiratory tract and heart

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 15

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 16
Figure 8.5: Derivatives of the mesoderm.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 17

Somite
Q. Write a short note on somite.
∙ Paraxial mesoderm on condensation form somitomeres
that later on segmentation forms somites from the
prechordal plate to the primitive streak.
∙ Proximal somitomers (underlying the developing brain,
rostral to the otic vesicle) remains unsegmented and do
not form somites. These forms striated muscles of face,
jaw and throat. (MCQ)
∙ Note: The prechordal plate mesoderm form preotic
somites that later forms extrinsic muscles of the eye.
∙ First pair of somites appears on the 20th day on each side
of the cranial end of notochord. (MCQ, Viva)
∙ Somites continue to appear from day 20 to day 30 in
cranio-caudal direction.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 18

Somite
∙ In human 42 to 44 pairs of somites forms and they can be
grouped as follows:
- Occipital 4 pairs
- Cervical 8 pairs
- Thoracic 12 pairs
- Sacral 5 pairs
- Coccygeal 8–10 pairs
∙ Many coccygeal pairs of somites eventually disappears till
it reaches final count of 37 pairs of somites.

Counting of somites

∙ First somite pair appears on day 20 and further three pairs

of somites get segmented each day until the end of the
fifth week. Hence, somites gives idea about the foetal
age. (MCQ, Viva)

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 19

Table 1: The correlation between embryo age with
number of somites. (MCQ)
Age in days Number of pairs of
(approximately) somites (approximately)
20 1–3
21 4–7
22 7–10
23 10–13
24 13–16
25 16–19
26 19–21
27 21–24
28 24–27
30 34–35
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 20
Structure of somite
∙ Somite is differentiated into three parts:
1. Sclerotome: It is ventromedial part and forms ribs and
vertebrae.
2. Myotome: It is middle part and forms skeletal muscles.
3. Dermatome: It is dorsolateral part and it forms the dermis
of the skin.
∙ A small cavity (myocele) of somite is obliterated by
proliferation of cells.
∙ Each somite is supplied by a single spinal nerve.
∙ Some authors consider dermomyotome instead of
separate dermatome and myotome.
∙ Note: The occipital somites and preotic somites participate
in the formation of base of skull.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 21

Box 1: Midline structures of trilaminar germ
disc
Q. Enlist the midline structures of germ disc.
∙ Trilaminar germ disc has following structures in the midline from cranial to caudal direction:
1. Septum transversum
2. Pericardial sac
3. Buccopharyngeal membrane
4. Notochord, neural tube
5. Primitive node
6. Primitive streak
7. Cloacal membrane

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 22

 Vasculogenesis

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 23

Box 2: Vasculogenesis and angiogenesis
∙ Vasculogenesis: Process of the formation of a new vessel.
∙ Angiogenesis: Sprouting of vessel into adjacent area by endothelial
budding.
∙ At the beginning of the third week, formation of vasculature begins in the
extra-embryonic mesoderm of the yolk sac, connecting stalk and chorion.
(MCQ)

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 24

Box 2: Vasculogenesis and angiogenesis
Steps of formation of vasculature

∙ On day 17, the mesenchymal cells differentiate into haemangioblasts.

∙ Haemangioblasts aggregate to form the cluster of cells called blood island.
∙ Haemangioblasts differentiate into two cell lineages as follows:
A. Primitive haematopoietic stem cells (HSC) – form erythropoietic cells, megakaryocytes and
primitive macrophages.
B. Endothelial precursor cells (EPC) – form epithelial lining of primitive endothelium.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 25

Box 2: Vasculogenesis and angiogenesis
∙ Small cavities appear in blood island to form lumen of primitive capillaries.
∙ Primitive haemangioblastic islands fuse to develop capillary networks (vasculogenesis).
∙ EPC sprout into adjacent mesenchyme and fuse with adjacent haemangioblastic island and
develop new capillaries (angiogenesis).
∙ By the end of third week, yolk sac, connecting stalk and chorionic villi get completely vascularised.
∙ Yolk sac is the first supplier of blood cells (up to 60 days). Later liver, spleen, thymus and bone
narrow take over the haematopoietic function.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 26

Folding of embryo
∙ Cells lying in the central part of the germ disc multiply rapidly than that at
the periphery. Overgrown central part result into the folding of germ disc
and forms primitive cylindrical embryo.
∙ Due to the overgrowing tissue, the embryo shows folding in the median
plane (cephalocaudal folding) and lateral plane (lateral folding).
∙ Cephalocaudal folding can be studied as cranial head fold and caudal tail
fold.
∙ Amniotic cavity enlarges to enclose ventral surface of the embryo and
amniotic membrane forms a tubular covering around the connecting stalk.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 27

Folding of embryo
∙ All the folds converge on the ventral surface of the embryo to produce
hour-glass constriction of the yolk sac.
∙ Part of yolk sac trapped within the embryo form primitive gut and the
protruding part of yolk sac (extra-embryonic part) forms umbilical
vesicle.
∙ Vitellointestinal duct temporarily connects primitive gut with umbilical
vesicle.
∙ Part of the primitive gut cranial to the vitellointestinal duct is foregut and
caudal to it is hindgut whereas the part of gut communicating with the
vitellointestinal duct is called midgut.
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 28
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 29
Head fold
∙ Overgrowing germ disc folds around cranial end of notochord to form
head fold.
∙ Foregut: Trapped part of yolk sac in the head fold.
Ventral relations of foregut: buccopharyngeal membrane,
pericardial sac and septum transversum.
Dorsal relations of foregut: notochord and brain vesicles.
Cranial relation of foregut: rapidly growing forebrain vesicle.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 30

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 31
Some interesting facts

∙ Due to the head fold, the buccopharyngeal membrane come to lie on the
ventral surface of embryo and becomes cranial to the pericardial sac and
septum transversum.
∙ During the fourth week, buccopharyngeal membrane ruptures.
∙ Foregut is separated from pericardial sac by cardiogenic plate
(mesenchyme that forms heart).
∙ Septum transversum is a mesenchymal tissue that lies caudal to the
pericardial sac. Septum transversum form fibrous pericardium, part of
diaphragm and part of ventral mesentery of the foregut.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 32

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 33
Tail fold
∙ Overgrowing germ disc also fold ventrally around
the caudal end of notochord to form tail fold.
The tail fold produces the following changes:
Hind gut: Trapped part of the yolk sac in the tail fold.
Dorsal relations: Notochord and neural tube
Caudal relations: Degenerating primitive node
and streak
Allantois: Communicates with hind gut
Bilaminar cloacal membrane: lies caudal to the
connecting stalk.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 34

Cloaca
∙ Caudal to the hindgut, a bilaminar area of the germ
disc is called cloacal membrane.
∙ At the site of cloacal membrane, ectodermal surface
shows a depression called ectodermal cloaca.
∙ Post-allantoic part of the hindgut dilates to form
endodermal cloaca.
∙ Cloacal membrane is divided into ventral urogenital
membrane and dorsal anal membrane.
∙ Both urogenital and anal membranes rupture in the
fifth week. (MCQ)

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 35

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 36
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 37
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 38
Lateral folds
∙ Overgrowing central part of the germ disc forms two
lateral folds (right and left).
Formation of lateral fold produces following
changes:
1. Amnioectodermal junction come to lie on umbilical
cord
2. Formation of midgut and vitellointestinal duct and
umbilical vesicle.
3. Formation of dorsal mesentery of gut:
Splanchnopleuric intra-embryonic mesodermal layer
form dorsal mesentery.
4. Peritoneal cavity: The somatopleuric intra-
embryonic mesoderm fuse ventrally in the midline to
convert intra-embryonic coelom into peritoneal
cavity.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 39

Box 3: Connecting stalk
∙ Formation of tail fold moves the attachment of connecting stalk form dorsal end of germ
disc to ventral aspect of embryo and limit it at the umbilical opening.
Structure of connecting stalk in the fifth week
1. Vitellointestinal duct
2. Remnants of the yolk sac
3. Extra-embryonic mesodermal connective tissue that later forms Wharton’s jelly
(connective tissue of umbilical cord) and vessels of umbilical cord
∙ Connecting stalk is the site of amnioectodermal reflection and continuation of intra-
embryonic and extra-embryonic coelom. This coelomic continuation persists up to 10 th
week.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 40

 Differentiation of
endoderm

Derivatives of
endoderm

∙ Flat endoderm — (embryonic

folding) → tube
∙ Endodermal tubular structure →
foregut, midgut and hindgut.
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 41
 Table 1: Derivatives of primitive gut (MCQ, Viva)
Part of the gut Derivatives
Foregut - Lining of epithelium of
Pharynx, oesophagus, stomach, duodenum up to the ampulla of the Vater
Tongue, floor of mouth
- Lining epithelium of respiratory system, auditory tube, tympanic cavity
- Parenchyma of tonsil, thyroid, parathyroid, thymus, liver and pancreas

Midgut - Lining epithelium of

Distal part of duodenum, jejunum, ileum, caecum, appendix, ascending colon, right two-third of
transverse colon
Hindgut - Lining epithelium
Left one-third of transverse colon, descending colon, sigmoid colon, rectum, anal canal up to
mucocutaneous junction
- Lining epithelium of
Urinary bladder, urethra, vagina
- Parenchyma of prostate, bulbourethral gland

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 42

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 43
Week wise changes in embryonic period
∙ Embryonic period extends from fourth week to eighth week.

∙ The changes of the embryonic period are listed below.

∙ At the beginning of fourth week, the embryonic disc is almost flat and has 4–12 somites.
Fourth week

• During the fourth week of the intrauterine life, following changes occurs:

∙ Increase in the somite numbers

∙ Formation of three brain vesicle from the neural tube

∙ Formation of head and tail folds

∙ Twenty-first days and later: Heart starts beating

∙ Twenty-fourth day: First pharyngeal arch appears (MCQ)

∙ Twenty-sixth day: Three pairs of pharyngeal arches

∙ 26th –27th day – rudimentary forelimb bud

∙ 28th day – rudimentary hind limb bud

∙ Formation of prominence of forebrain vesicle

∙ Otic pits, lens placodes (ectodermal thickening) appears

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 44

Fifth week
• During the fifth week of the intrauterine life, following changes occurs:
∙ Rapid growth of facial and head prominences
∙ Rapid growth of a second pharyngeal arch
∙ Growth of the limb buds
∙ Formation of alar and basal laminae of the neural tube
∙ Appearance of olfactory placodes, maxillary and frontonasal process.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 45

• Sixth week

• During the sixth week of the intra-uterine life, following changes occur:

∙ Shows spontaneous movements

∙ Growth of brain vesicles
∙ Appearance of nasal processes
∙ Buccopharyngeal membrane ruptures
∙ Development of appendix, caecum and spleen
∙ Differentiation of the limb digits begins. Differentation in lower limb starts 4–5 later than upper limb.
∙ Umbilical herniation is common.

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 46

• Seventh and eight weeks

• During the seventh and eight weeks of the intra-uterine life, following changes occur:

∙ Development of face, external ear (auricular hillock) and eye

∙ Well defined limbs and digits
∙ Development of metanephric kidney
∙ Development of testes and ovaries
∙ Differentiation of external genitalia
∙ During eighth week, coordinated limb movements occur
∙ Primary ossification centres start appearing
∙ Caudal tail-like eminence disappears

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 47

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 48
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 49
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 50
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 51
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 52
Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 53
Thank you………….

Human Embryology/Yogesh Sontakke/2nd edn/CBS Publishers 54