The in-ovo and in-vitro observation of

chicken embryo development

D. Listyorini, D.Sc.
Animal Tissue Culture Laboratory
Department of Biology
State University of Malang

D.Listyorini.chick.embr.dev.
Introduction
Chicken embryo develops in the shell composed of
mostly calcium.
Since the embryo develops outside the body of the
mother, the shell is an important protection against
harmful conditions.
Beside, the shell is the main source of calcium which is
necessary for the development of skeleton and cell
junctions.

D.Listyorini.chick.embr.dev
Introduction
Chicken embryo is one of good models for
studying vertebrate embryo development,
because we can observe its development
directly time by time.
We can study chicken embryo development
through in-ovo or in-vitro observations.

D.Listyorini.chick.embr.dev
Method 1: In-ovo observation
Tools:
Small glass bowls
Petri disc
Shell cutter
Cellophane tape
Dry oven

Shell cutter
Cellophane tape

D.Listyorini.chick.embr.dev
Method 1: In-ovo observation
Procedure:
Lay the eggs horizontally to let the yolk
stabilizes the position of the embryo.
Draw a mark to point the upper part of egg.

D.Listyorini.chick.embr.dev
Method 1: In-ovo observation
After5 to 10 minutes, swap the shell carefully
with 70% Ethanol. Make sure the position of
egg does not change (do not turn the egg!)

X Up

X
Down

D.Listyorini.chick.embr.dev
Method 1: In-ovo observation
Put back on the pedestal.
Pinch a small hole on the
blunt end of the egg to
decrease the internal
pressure. Small Cut
hole
Cutthe shell carefully
using shell cutter. pedestal

D.Listyorini.chick.embr.dev
Method 1: In-ovo observation
Cellophane
Up
Discard the cut shell. tape

Cover the holes with cellophane

tape.
Putthe egg in the pre-sterilized Down
glass bowl (with 70% Ethanol).
Cover the bowl with petri disc.
Incubatein the 380C dry
incubator.
Puta cup of water in the
incubator to maintain the
humidity. D.Listyorini.chick.embr.dev
Method 2: In-vitro observation
Tools:
Small glass bowls
Petri disc
Shell cutter
Dry oven

Shell cutter
D.Listyorini.chick.embr.dev
Method 2: In-vitro observation
Procedure:
Lay the eggs horizontally to let the embryo on
the upper part of yolk to stabilize the position
of the embryo.
Draw a mark to point the upper part of egg.

pedestal
D.Listyorini.chick.embr.dev
Method 2: In-vitro observation
Sterilize glass bowl with 70%
Ethanol. Dry carefully!
After 5 to 10 minutes cut the Embryonic
egg shell carefully, keep upper disc
part up! Albumen
Pour albumen and yolk
carefully in the bowl!
Be careful, not to tear yolk sac! Yolk
Make sure the embryonic disc
is located at the upper part of
yolk.

D.Listyorini.chick.embr.dev
Method 2: In-vitro observation
Cover the bowl with
petri disc.
Incubate in the 380C
dry incubator.
Put a cup of water in
the incubator to
maintain the humidity.

D.Listyorini.chick.embr.dev
Method 2: In-vitro observation
Do the observation on the embryo
development every 24 hours.
Pay attention on the increase of
embryonic disc size, the development of
blood vessel on the yolk, the development
of the chicken body and specific structures
of chicken.

D.Listyorini.chick.embr.dev
Result: In-ovo observation
3 day 4 day
Embryonic disc Embryo

8 day Death Embryo

Yolk

Yolk
Embryo

Blood vessel
D.Listyorini.chick.embr.dev
Result: In-vivo observation
2 day 3 day
embryo embryo

yolk yolk

Blood Blood
vessel vessel

4 day 5 day embryo

embryo
yolk
yolk

Blood Blood
vessel vessel

D.Listyorini.chick.embr.dev
Result: In-vivo observation
6 day 7 day

8 day 9 day

D.Listyorini.chick.embr.dev
Result: In-vivo observation
10 day 13 day

14 day 16 day

D.Listyorini.chick.embr.dev
Result: In-vivo observation
17 day 18 day

19 day 20 day

D.Listyorini.chick.embr.dev
 Result: In-vivo observation
21 day 21 day

D.Listyorini.chick.embr.dev
Conclusion
It is possible to observe the development of chicken
embryo directly.
In-vitro maintenance allowed chicken embryo to develop
until 21 day.
Chicken showed malformation in appendages and body
wall development.
Malformation is suggested to be caused by the lack of
the shell as the source of calcium.

D.Listyorini.chick.embr.dev