INTRODUCTION HISTORY CELL CLONING TYPES OF CELL CLONING TECHNIQUES OF CELL CLONING APLLICATIONS CONCLUSION REFERENCES

1. TECHNIQUES OF CELL CLONING
By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )

2. SYNOPSIS
INTRODUCTION
HISTORY
CELL CLONING
TYPES OF CELL CLONING
TECHNIQUES OF CELL CLONING
APLLICATIONS
CONCLUSION
REFERENCES

3. A population of identical molecules
(genes), cells or organisms, all of which
are derived from the same parent by
asexual means, is known as a clone.
INTRODUCTION

4. History
1963:-J.B.S. Haldane coins the term ‘clone’.
1978-The first baby conceived through in
vitro fertilization is born in England.
1994-Dr. Neal First of the University of
Wisconsin clones calves from embryos that
had grown to 120 cells.
1996-Dolly is born at the Roslin Institute.

5. What is cloning
The process of making identical genomic copies
of an origanism
Production of one or more individual plants or
animals(whole/in part) that are genetically
identical

6. Types of Cloning
1. Embryo Cloning
2. Reproductive cloning
3. Therapeutic Cloning

7. a) Embryo cloning
Produce mono zygotic twins or triplets
Fertilized embryo
remove
one/more cells
Encourage to develop into embryo
Twins /triplets have identical DNA

10. b) Reproductive cloning
Produce a duplicate of existing animal
Used to clone sheep & other mammals
Produce several genetic defects

11. Newly formed embryo containing DNA from somatic cell
cell division
implant
(Diploid )

13. c) Therapeutic Cloning
Somatic Cell Nuclear Transfer(SCNT)
Biomedical Cloning
Research Cloning
Regenerative Medicine
Nuclear Transplantation Therapy(NTT)

14.  Therapeutic cloning (research cloning) is when stem cells are
extracted to grow into a piece of human tissue which is
encouraged to grow into a human organ for transplant

15. Therapeutic Cloning
DNA is extracted from a human’s cell
inserted into a woman’s ovum
develop and produce stem cells.
stem cells are removed from the pre-embryo
grown into specific organ
transplanted into the patient.

16. Cloning Techniques
Somatic Cell Nuclear Transfer-
In this process, the nucleus of a somatic cell is
removed and inserted into an unfertilized egg .
The Roslin Technique
The Roslin Technique is a variation of somatic cell
nuclear transfer that was developed by researchers a
the Roslin Institute. The researchers used this
method to create Dolly.
The Honolulu Technique
The Honolulu Technique was developed by Dr.
Teruhiko Wakayama at the University of Hawaii. In
this method, the nucleus from a somatic cell is
removed and injected into an egg that has had its
nucleus removed.

17. Dolly the Sheep
Dolly, a Finn-Dorset ewe, was the first mammal to have been
successfully cloned from an adult cell.
She was cloned at the Roslin Institute in Scotland and lived there
from her birth in 1996 until her death in 2003 when she was six.

18. Cloning may be carried out in two approaches
:-
 Monolayer culture:
 Petri dishes, multiwell plates or flasks can be
used for cloning by monolayer culture.
 Suspension culture:
 cloning can be carried out in suspension by
seeding cells into viscous solutions
(methocel) or gels (agar).

19. Dilution Cloning

20. Suspension cloning technique in agar
cell cloning
Cells from suspension culture or
monolayer can be used
Monolayer cells have to trypsinized
while suspension cells can be
directly used
Count and dilute the cells serially
Freshly prepared agar medium with
appropriate dilution is used.
Agar medium is inoculated with the
diluted cells and incubate for 1-3
weeks.

21. Conditions That Improve Clonal Growth or culture factors:-
 Medium.
 Choose a rich medium, such as Ham’s F12, or a medium that
has been optimized for the cell type in use,
 Serum.
 When serum is required, fetal bovine is generally better than
calf or horse.
 Hormones.
 Insulin,has been found to increase the plating efficiency of
several cell types
 Dexamethasone, a soluble synthetic hydrocortisone analogue,
improves the plating efficiency of chick myoblasts
fibroblasts,and melanoma and gives increased clonal growth.

22. Intermediary metabolites.
 Supplementing the medium with intermediary metabolites (e.g,
pyruvate, α- ketoglutarate and nucleosides stimulates plating
efficiency.
 Carbon dioxide.
 CO2 is essential for obtaining maximum cloning efficiency for
most cells. Although 5% CO2 is usually used, 2% is sufficient
for many cells.
 Treatment of substrate.
 Polylysine and Fibronectin also improves the plating of many
cells
 Trypsin.
 Purified trypsin used at 0.05 μg/mL

23. Why need Animal Cloning
Research?
 to generate groups of genetically identical
animals for research purposes
 to rapidly propagate desirable animal stocks
 to improve the efficiency of generating and
propagating transgenic livestock
 to produce targeted genetic alterations in
domestic animals
 to pursue basic knowledge about cell
differentiation

24. Application of cell cloning
 There are many applications that nuclear
transfer cloning might have for biotechnology,
livestock production, and new medical approaches.
 Work with embryonic stem cells and genetic
manipulation of early embryos in animal species
(including nuclear transfer) is already providing
unparalleled insights into fundamental biological
processes and promises to provide great practical
benefit in terms of improved livestock, improved
means of producing pharmaceutical proteins, and
prospects for regeneration and repair of human
tissues.
 In agricultural industry
 Drug production

25. LIMITATIONS
1. Developing cures
2. Is an embryo a human person ?
3. Stability of stem cells
4. Where would the eggs come from ?
5. Health risk concerning the donors
6. Genetic make-up is altered
7. Mutants are created
8. Clones are unhealthy
9. Plant cloning has been around for thousands of
years
10. animal cloning has been around for over 20 years

26. Conclusion
The term “clone” has many meanings but
in its simplest and most scientific sense it
means the making of identical copies of
molecules, cells, tissues, and even entire
animals

27. References
Books :
R.Ian Freshney-culture of animal cell
U.satayanarayana-biotechnology