Rajesh Wakchaure et al Int. J. Phar. & Biomedi. Rese. 2015, 2 (5): 11-16 ISSN: 2394 – 3726
Copyright © 2015; IJPBR 11
A Review on Cryopreservation of Embryos and its Research Implications in
Animal Breeding and Reproduction
Rajesh Wakchaure
1
, Subha Ganguly
2
*, Subhash Sharma
3
, Praveen Kumar Praveen
4
, Maneesh
Sharma
5
and Tanvi Mahajan
6
1
Dept. of Animal Genetics & Breeding,
2
Dept. of Veterinary Microbiology,
3
Dept. of Veterinary Parasitology,
4
Dept. of Veterinary Public Health & Epidemiology,
5
Dept. of Veterinary Pathology,
6
Dept. of Veterinary Anatomy & Histology, Arawali Veterinary College
(Affiliated with Rajasthan University of Veterinary and Animal Sciences, Bikaner)
N.H. – 11 Jaipur Road, V.P.O. Bajor, Dist. Sikar, Pin – 332001, Rajasthan, India
*Corresponding Author E-mail: ganguly38@gmail.com
INTRODUCTION
Embryo cryopreservation has been a useful tool for embryology since the early 1970s. It has become an
essential part of assisted reproductive technologies, allowing long term storage of valuable embryos from
lab animals, livestock, endangered species and humans. Initially, work was done with conventional slow
cooling technologies using cryoprotectants such as glycerol or ethylene glycol and sucrose. More
recently, however, a greater advance has been made with vitrification, a procedure that bypasses ice
crystal formation and places the embryo in a glass-like state. This technology is receiving greater
attention as it is simpler, faster and less expensive, and provides the embryo with greater protection from
cryoinjury.
Principle of Embryo Cryopreservation
For prolong Storage of embryos, one has to attain a suspended stage of animation where cleavage of
embryo would be suspended and cellular activities would be practically at standstill. The safest method to
achieve this goal is to expose the embryos to progressively lower temperature and finally store in liquid
nitrogen (-196
o
C) or liquid helium (-296
o
C).
There are six step storage of embryo to successful embryo cryopreservation i) exposure to cryoprotectant,
ii) cooling to subzero temperatures, iii) storage, iv) thawing or warming, v) removal of cryoprotectant and
vi) return to a physiological environment [1-3]. Two important parameters determine the success of all
cryopreservation protocols 1) Rate of cells regaining equilibrium in response to cooling 2) Speed of
freezing [1-3]. Cryoinjury can occur from formation of intracellular or extracellular ice crystals, chemical
toxicity and osmotic injury [4] and is dependent upon size and shape of cells, membrane permeability,
Available online at www.ijpbr.org
ISSN: 2394 - 3726
Int. J. Phar. & Biomedi. Rese. 2015, 2 (5): 11-16
ABSTRACT
The goal of embryo cryopreservation is long term storage and reproducible high survival rates of
embryos following warming, leading to the successful establishment of pregnancy and live offspring
following embryo transfer. Successful cryopreservation requires a good understanding of the proper
use of cryoprotectants, sugars and macromolecules in order to allow the highest survival rates and
ultimately pregnancy and live offspring following embryo transfer. The success is dependent upon
many variables, including embryo stage and quality, embryo species, cooling and warming rates and
culture conditions. Efforts to optimize these conditions will further enhance survival and future
developmental potential of the embryo.
Key words: Cryoprotectants, Embryo cryopreservation, Reproduction, Vitrification