New Technologies to Produce Baby - PowerPoint Presentation

Slide1

New Technologies to Produce Baby

Slide2

Assissted Reproductive Technology (ART)

Artificial insemination (AI)

Male infertility factors

Inject the sperm directly in the female reproductive tract (why)

Artificial insemination is intra-vaginal or intra- cervical injection/transfer of semen to facilitate fertilization

Slide3

AIH

: Artificial insemination using husband’s semen

AID: Artificial insemination using donor’s semen

IUI: Intra-uterine insemination

SIFT

: Sperm intra-fallopian transfer

DIPI: Direct intra-

peritonial

insemination

Slide4

Artificial Insemination using husband’s Semen (AIH)

Oligospermia

Anatomical defects in male reproductive system

Centrifugation-concentration of sperms in the semen

Accumulation using deep freeze storage

Slide5

Split Ejaculate Method

During ejaculation the semen comes in two or three fractions

The first half ejaculate contains majority of spermatozoa; also the sperm quality in terms of motility and morphology is better than the one ejaculated in later half

Substances such as

kalikrein

,

arginine

or albumin improve motility of sperms in the split ejaculate

Cleaning of ejaculate to obtain large number of live and motile sperms

Slide6

Artificial insemination using donor’s semen (AID)

Husband

Azoospermic

or

nacrospermic

or

asthenospermic

or severely

oligospermic

Donor sperms- sperm banks

Collection, storage by freezing the sperms from anonymous donors

Strict guidelines and regulatory standards for donor selection, cryopreservation and semen banking

Confidentiality

Testing of semen for any microbiological contamination

Slide7

Artificial Insemination by Donor Sperm AID

Many women use donor sperm with the artificial insemination procedure

If male partner’s sperm is of poor quality or quantity

Some men with genetic disorders may choose AID so they don’t pass on genetic disorders to their children

Using a sperm donor is also a choice for single women or lesbian couples

Before proceeding with donor sperm and the artificial insemination procedure, doctor may

encourage

to speak to a counselor about any anxieties or concerns.

Slide8

Procedure of Ovarian hyper-stimulation

The aim of controlled ovarian hyper-stimulation is to correct subtle cycle disorders, to increase the number of available oocytes for fertilization and to improve the timing of insemination

Slide9

Procedure of Ovarian hyper-stimulation

The two most frequently used drugs are

clomiphene

citrate and

gonadotropins

Clomiphene

citrate blocks the estrogen receptors of the hypothalamus

This altered feedback information causes the hypothalamus to make and release more

gonadotropin

releasing hormone (

GnRH

)

This in turn causes the pituitary to make and release more FSH and LH

Slide10

Gonadotropins

The

gonadotropins

Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH) are released under the control of

gonadotropin

-releasing hormone (

GnRH

) produced in the hypothalamus.

FSH directly affects the production and maturation of follicles by stimulating the

granulosa

cells of ovarian follicles

LH stimulates the

thecal

cells to produce testosterone (and indirectly

estradiol

)

Slide11

Gonadotropins

LH surge is obligatory for the rupture of the dominant follicle

The first FSH preparation used in fertility practice was extracted from postmenopausal urine (human menopausal

gonadotropin

,

hMG

)

In the 1990s, recombinant DNA technology led to the development and clinical introduction of human recombinant FSH (

rFSH

).

Slide12

Monitoring, Timing, and/or Induction

of Ovulation

In IUI cycles with ovarian

hyperstimulation

the follicular growth is always monitored by

sonography

, since uncontrolled

multifollicular

growth may lead to (high order) multiple pregnancies

For the detection of ovulation, either LH-surge tests can be used or ovulation can be induced by human chorionic

gonadotropin

(

hCG

).

Slide13

Intra-uterine Insemination

Most common method of artificial insemination

It makes the trip shorter for the sperm and gets around any obstructions

The

sperms escape the unreceptive cervical

mucus

Slide14

IUI

The success of IUI is based on three steps

Firstly, semen is processed, so that motile spermatozoa are concentrated in a small volume and the number of motile sperms is increased at the site of fertilization

Secondly, the insemination takes place directly, bypassing the possibly hostile cervical mucus and bringing the semen in closer proximity of the oocyte

Thirdly, optimizing the timing by monitoring or inducing ovulation

All these steps increase the probability of

conception, especially

in the case of compromised semen parameters

Slide15

 Preparing the Sperm

In IUI the sperm sample undergoes a special washing and processing procedure

After a semen sample is obtained, the sperm are washed and concentrated

A semen sample is washed in the lab to separate the sperms from the seminal fluid

During this washing and processing phase, potentially toxic chemicals are removed, along with a seminal plasma that surrounds each sperm cell

Slide16

Under normal circumstances, sperm must pass through the cervical mucus to reach the uterus, a process that limits uterine entrance to only the healthiest sperm and prevents dead sperm, debris, and seminal fluid from getting in

Due to the presence of prostaglandins and microbes in the seminal fluid, semen cannot be placed directly into the uterus without the potential of inducing painful cramping and infection

During sperm preparation seminal fluids,

nonmotile

cells, and debris are removed, sperm can be placed directly into the uterus

Slide17

Besides removing prostaglandins, processing semen samples prior to IUI has other advantages

Sperm from semen samples can be concentrated and reduced to a volume that is consistent with what the uterine cavity can hold

Decapacitation

factors present in the semen are removed, increasing the sperm’s fertilization ability

Processing semen samples can also select for fractions of sperm with the best motility and improve overall motility

The prepared sample of sperm can be placed in preparation media known to support sperm

capacitation

and survival

Slide18

Basic Wash

The basic wash is the simplest and least expensive sperm preparation to perform. It is able to concentrate the sperm into a workable volume for insemination, removing the seminal fluid with its associated

decapacitation

factors and prostaglandins

Besides its simplicity, the advantage of a basic wash is that essentially all of the sperm in the ejaculate will be recovered

This is especially useful for samples with a low concentration of motile sperm. It is also beneficial for

cryopreserved

samples, or other fragile samples that would benefit from reduced processing and increased recovery

Slide19

Basic Wash

The disadvantage of a basic wash is that it concentrates all of the live sperm, dead sperm, white blood cells, red blood cells and seminal debris together. As a result, the healthy sperm are exposed to higher levels of reactive oxygen species (ROS), which can decrease sperm function and negatively affect fertilization

Therefore, the basic wash is not recommended for regular samples when there are a lot of cellular contaminants or dead cells present

Slide20

Density Gradient Preparation

The density gradient preparation consists of filtering sperm by centrifugal force through either one or multiple layers of increasingly concentrated

silane

-coated silica particles

Typically, semen is placed directly on top of the density gradient layers. During centrifugation, the most motile sperm pass through the different layers, making a soft pellet at the bottom.

The seminal fluids, dead or

nonmotile

sperm, and cellular debris are held up at the interfaces between the layers

Slide21

Density Gradient Preparation

The sperm that have successfully

pelleted

at the bottom can then be recovered, washed to remove the density gradient media, and used for IUI.

The ability to select for a population of clean, motile sperm makes it very useful for samples with a lot of dead sperm, round cells, or debris

Unless it is necessary to remove WBCs, it is not advisable to use density gradient preparations for samples with low concentrations of progressively motile sperm, as recovery may be limited

Slide22

Preparation of

Cryopreserved

Sperm

Patient Samples

In many instances, patients choose to use

cryopreserved

sperm that was stored prior to chemotherapy or military duty, or for other personal reasons.

As

cryopreserved

sperm are less robust than those in fresh samples, extra precaution should be taken when they are prepared

Generally,

cryopreserved

samples should not be subjected to more than a single wash in sperm wash media

Slide23

Preparation of

Cryopreserved

Sperm

The centrifugation time should also be reduced to 6–8 min

Patients should be educated that

cryopreserved

samples will have reduced sperm motility compared to the fresh state of the sample before freezing

Depending on the circumstances and the number of frozen vials a patient has stored, a plan should be made to thaw a sufficient number of vials to give the patients a reasonable chance of achieving pregnancy.

Slide24

Preparation of

Cryopreserved

Sperm

Donor Samples

Many patients now choose to take advantage of the easy access to sperm donor profiles provided by the Internet and order donor sperm from sperm banks across the country

The sperm samples are typically shipped just prior to their anticipated use

If the sample is not going to be used right away, measures should be taken to place it in a more permanent storage container

Slide25

Preparation of

Cryopreserved

Sperm

Most sperm banks are now providing donor samples that are prewashed. Under these circumstances, all that is necessary is to thaw the vial, check a drop of the warmed sample to confirm if it meets the sperm bank’s standards, and load it into the insemination syringe

If the sample was not washed prior to freezing, it will be necessary to wash the sample using the simple wash procedure, shortened to 6–8 min of centrifugation, prior to intrauterine insemination

Slide26

The IUI procedure is performed around the time of ovulation, typically about 24-36 hours after the surge in LH hormone that indicates ovulation will occur soon

A catheter is used to insert the sperm directly into the uterus

This process maximizes the number of sperm cells that are placed in the uterus, thus increasing the possibility of conception

Slide27

IUI- intra-uterine insemination

Slide28

Slide29

Modalities of Insemination

IUI is commonly performed between 36 and 42 h after a positive LH surge test

IUI 33 or

39h

after the administration of

hCG

resulted in the same pregnancy results, and therefore the timing of the insemination may depend on local convenience of the hospital

Insemination has to be performed only once per cycle

A 10-min interval of bed rest after IUI has a positive effect on the pregnancy

Slide30

Intra-tubal insemination

Intra-tubal insemination (ITI) involves injection of washed sperm into the fallopian tube, although this procedure is no longer generally regarded as having any beneficial effect compared with IUI 

ITI is different from gamete intra-fallopian transfer, where both eggs and sperm are mixed outside the woman's body and then immediately inserted into the fallopian tube where fertilization takes place.

Slide31

Direct intraperitoneal

insemination (DIPI)

The sperms are injected into the lower abdomen through a surgical hole or incision, with the intention of letting them find the oocyte at the ovary or after entering the genital tract through the 

ostium

of the fallopian tube

Slide32

Intra-uterine

Tuboperitoneal

Insemination (IUTPI)

Intrauterine

tuboperitoneal

insemination (IUTPI) involves injection of washed sperm into both the uterus and fallopian tubes

The cervix is then clamped to prevent leakage to the

vagina

The

sperms are

mixed to create a volume of 10 ml, sufficient to fill the uterine cavity pass through the interstitial part of the tubes and the 

ampulla

, finally reaching the peritoneal cavity and the Pouch of

Douglas

Slide33

Intra-uterine

Tuboperitoneal

Insemination (IUTPI)

IUTPI

can be useful in unexplained infertility, mild or moderate male infertility, and mild or moderate endometriosis

In non-tubal sub fertility, fallopian tube sperm perfusion may be the preferred technique over intrauterine insemination