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Reproductive Biotechnology - Coggle Diagram
Reproductive
Biotechnology
Ovum pick up
consists in the transvaginal removal of the oocytes by aspiration of the ovarian follicles with the aid of an ultrasound probe
absolutely harmless to the donor, the time taken for the collection is 15-20 minutes during which the donor is contained in a cattle crush.
Method
Transvaginal ultrasound probe placement
Aspiration of ovarian follicles
The oocytes collected from the ovarian follicles are selected in the laboratory and transferred to an incubator in a suitable culture medium to complete the maturation phase.
In vitro Fertilisation
a process of fertilisation in which an oocyte is combined with a sperm outside the female reproductive apparatus but in "in glass” in a lab
METHODS
Conventional insemination
Ovum pick-up when the oocyte is mature
semen preparation
Eggs are placed on a culture dish and surrounded by a number of spermatozoa ranging between 100,000 and 500,000 for a period of no less than 12 hours.
Spontaneous fertilization is expected.
Intracytoplasmic sperm injection
Ovum pick-up when the oocyte is mature
semen preparation
consists of the insertion of a selected sperm into each mature egg via microneedle injection.
Sperm Sexing
Tecnique: flow cytometry separation of spermatozoa based on the amount of DNA contained in the sperm
is based on the staining of the sperm cells with
a fluorescent dye and sorting according to the DNA content
advantanges
the use of sexed semen is an essential solution for the animals with sex-linked diseases and it helps in reducing costs related to unwanted male or female specimens within specific animal supply chain.
disadvantanges
Reduced fertiliy (10-20% compared to conventional semen)
More expensive
Embryo sexing
based on the chromosomal difference that existsbetween embryos of the two sexes.
The diagnosis of sex is 97% accurate.
The presence or absence of the Y chromosome is the
criteria adopted to sort embryos.
fasi
biopsy is used, i.e. the removal of some cells of the embryo using
a micro blade, in order to obtain a:
DNA sample to be analysed . This DNA is multiplied through a
procedure called
PCR (Polymerase Chain Reaction) which can help to highlight the
presence or absence of the Y male chromosome
electrophoresis of DNA samples in an agarose gel
spectrophotometric analysis of the DNA
Embryo splitting
obtain two (or more) genetically identical individuals from the
same embryo, taken from the same female donor (same mother)
METHOD
Transvaginal ultrasound probe placement
Embryo collection
Microsurgical blade cut
Obtainment of two identical embryos
Immediate transfer to synchronized recipient cows or freezing
Nuclear Tranplantation
to obtain two (or more) genetically identical animals
from a somatic cell taken from a female donor.
It consists in replacing the genome of one cell with the one derived from the other one.
TECHNIQUE
Transvaginal positioning of the ultrasound probe
the nucleus is extracted from a cell with a pipette and introduced into a previously enucleated oocyte.
special solutions are used in order to stimulate the oocyte
to divide and give rise to an embryo. The sequence of
events taking place are technically defined as oocyte
activation”.