Ppt

1. BY
DR.NAZIA SHAHZADI

2. WHAT IS IN-VITRO
FERTILIZATION
 A medical procedure whereby an egg
is fertilized by sperm in a test tube or
elsewhere outside the body.
 The process involves monitoring and
stimulating a female's ovulatory process,
removing an ovum or ova from their
ovaries and letting sperm fertilise them
in a culture medium in a laboratory

3. IVF STEP-BY-STEP
 Step 1: Control Ovarian Hyperstimulation
(COH) COH is done using different
protocols. ...
 Step 2: Egg Retrieval. Oocytes. ...
 Step 3: Fertilization and Embryo Culture.
2PN. ...
 Step 4: Embryo Quality. There are several
criteria used to assess the quality of the
embryo. ...
 Step 5: Embryo Transfer.

4. TYPES OF IVF
The three main approaches to IVF that
involve no or fewer drugs are
 Natural cycle IVF:- Natural IVF works
within a woman's natural cycle. It is
significantly more gentle on the body
than conventional IVF.
 Mild stimulation IVF:- With mild
stimulation IVF, you receive a lower
dose of fertility drugs over a shorter
period of time than with standard IVF.

5. CONTINUED
 That reduces your treatment time by about
two weeks and means you avoid a lot of
the unpleasant side effects from the drugs.
 In vitro maturation (IVM):- In vitro
maturation (IVM) is when a woman's eggs
are collected and matured outside the
body. This is done as part of an in vitro
fertilization (IVF) procedure. A woman's
eggs (also called oocytes) are formed
before she is born.

6. WHAT IS THE LATEST
TECHNIQUE OF IVF?
 The most advanced IVF procedure
is laser-assisted hatching which is useful
in certain conditions. It entails
developing a hole in the embryo's tough
exterior shell, so that it becomes easier
for in vitro embryo to implant.

7. SUCCESS RATE OF IVF
The Society for Reproductive Technology
(SART) states that
 for women under 35, the percentage of
live births via IVF is 55.6%.
 Live births per first embryo transfer is
41.4%.
 With a later embryo transfer, the live
births percentage is around 47%.

8. CONTINUED
 In 2019, the percentage of IVF
treatments that resulted in a live birth
was:
 32% for women under 35
 25% for women aged 35 to 37
 19% for women aged 38 to 39
 11% for women aged 40 to 42
 5% for women aged 43 to 44
 4% for women aged over 44.

9. CHEMICAL MEDIATORS
OF IVF
 CYTOKINES
 HORMONES
 PROTEINS

10. CYTOKINES
 Cytokines are the major immune regulators
in pregnancy, mediating gametogenesis,
uterine receptivity, implantation reactions,
embryogenesis and fetal development and
the onset of parturition either by triggering
inflammatory processes or by inducing
immune tolerance.
 Elevated cytokine ratios have been linked
with adverse reproductive outcomes.

11.  Many miscarriages are caused by or
associated with abnormal levels of
cytokines, which are proteins secreted by
white blood cells to control the
inflammatory process in the body. The
activity of these cytokines can be broken
down into two categories: pro-inflammatory
(or TH1) and anti-inflammatory (TH2).
 The capacity to produce monomeric
cytokines, either pro-inflammatory, (TNF-α,
IFN-γ) or anti-inflammatory (IL-10), exists
in many lymphocyte subsets.

12. Follicular cytokines
(proinflammatory):-
 IL-1β:- Evidenced to participate in
ovulation induction by facilitating follicular
rupture
 IL-6:- stimulation of cell proliferation,
differentiation, and survival—all essential
for blastocyst development and
implantation.
 IL-18:- Indirectly supports the role of IL-18
in follicle maturation.
 IFN-γ:- to overcome apoptosis inhibition &
to decrease cell proliferation in
endometriosis.

13. CONTINUED:-
 IFN-α:- positive correlation between
follicular IFN-α levels and follicular
diameter from pre-ovulatory granulosa
cells.
 TNF-α:- Deteriorate the microenvironment
in the follicle, thereby negatively affecting
oocyte and embryo quality.
 IL-12:- Associated with a negative effect on
folliculogenesis, oocyte quality and
implantation
 IL-23:- participate by promoting
inflammation, a hallmark of endometriosis.

14. ANTI-INFLAMMATORY
CYTOKINES
G-CSF:- Maintaining healthy endometrium ( successful
implantation & furthur development of embryo).
 Useful biomarker of oocyte competence before
fertilization.
 Improve implantation rate & successful pregnancy
outcome in IVF.
M-CSF:- Levels in serum may reflect successful
stimulation and ample follicle maturation.
Once the blastocyst has traversed the basal membrane,
the migration of first trimester invasive trophoblast in
vitro requires the expression of alpha 5 and beta 1,
integrins.
IL-10:-

15. CHEMOKINES
 MIP-1α:- MIP-1α in PCOS patients may
reflect a character of increased inflammation in
stressed ovaries.
 MIP-1β:- to promote folliculogenesis and
pregnancy establishment.
 MCP-1:- MCP-1 is a potent chemo attractant
of monocytes and T lymphocytes.
 RANTES:- RANTES are potent chemo
attractant of monocytes and T lymphocytes.
 IL-8

16. BIOMARKERS
 sAPO-1/Fas:- sAPO-1/Fas mediates
apoptosis inhibition, which is important
in preventing oocyte from succumbing to
atresia during follicular maturation.
 CD44(v6):- macrophage membrane-
expressed CD44 protein has been
shown to participates in clearance of
apoptotic granulosa cells.

17. Analysis of cytokines
Cytokine origin was studied by mRNA analysis
of granulosa cells using Real time PCR.
 Higher follicular MIP-1α and CD44(v6) were
found to correlate with polycystic ovary
syndrome,
 IL-23, INF-γ, and TNF-α with endometriosis,
 Higher CD44(v6) but lower IL-β and INF-
α correlated with tubal factor infertility
 the TGF-beta super family:- Help in decidual
reaction

18. CONTINUED
 Lower levels of IL-18 and CD44(v6) characterized
unexplained infertility.
 IL-12 positively correlated with Oocytes fertilization
and embryo development,
 while increased IL-18, IL-8, and MIP-1β were
associated with successful IVF-induced pregnancy.
 Invasion is mediated by several classes of
proteinases including serine proteases,
metalloproteinase and collagenases that degrade the
extracellular matrix (ECM).
 The insulin-like growth factor (IGF) system has been
shown to be involved strongly in endometrial
proliferation and differentiation.

20. HORMONES:-
 Hormones like
 AMH,
 LH,
 FSH,
 progesterone,
 estrogen,
 prolactin
 TSH
 Beta HCG play a huge role in female
reproductive health and hence, the process of
IVF.

21. AMH (Anti-mullerian hormone)
 an AMH test at the beginning of IVF
journey should be performed. Since AHM
starts declining in women after the age of
25, it’s important to know reproductive
lifespan, and AHM can be of great help for
this.
 So, blood is taken from vein to determine
the level of AMH in the body, which can be
done at any point of periods. As a result,
high (>1.0 ng/mL) or low (<1.0 ng/mL) level
of AHM obtained showing a high or low
ovarian reserve, respectively.

22. LH (Luteinizing hormone)
 LH provokes the creation of the corpus
luteum, which is a structure in the female’s
ovaries. This structure stimulates the
production of progesterone and supports
pregnancy at its beginning.
 Levels of LH in the body may increase with
age, so it’s important to know whether the
level of this hormone is normal in the body
when thinking of starting IVF. This means
to take an LH test, which requires a blood
sample.

23. RESULT OF LH TEST:-

24. FSH (Follicle-stimulating
hormone)
 Follicle-stimulating hormone (FSH) is
the growth hormone responsible for
producing mature eggs in the ovaries. It
is also the hormone that is inject able
and used in the IVF cycle for infertility
treatment.
 The level of FSH in a woman’s body is
constantly increasing with age, even
after menopause when there are no
eggs left.

25. FSH CONTINUED:-
 In the process of IVF, an FSH test is conducted on
the 3rd day of menstrual cycle to determine whether
baseline level of the hormone is normal (>9). In case
it is increased (9-20), this means that ovarian
reserve is low. The level of FSH can also predict
body’s reaction to fertility medication.
 When the level of FSH on day 3 of menstrual cycle is
higher than 20, there is an extremely low chance that
body will react to ovarian stimulation via fertility
drugs. However, each patient is approached
individually, so it’s best to consult with fertility doctor
before jumping to any conclusions.

26. Progesterone
 This hormone’s role is to prepare the uterine lining,
the endometrium, for pregnancy in case it occurs. If it
doesn’t, the level of progesterone decreases, and
the period starts.
 In fact, this hormone takes care of the lining’s ability
to allow the egg to stick to it when it gets fertilized. In
case of pregnancy, progesterone may keep being
produced for 8-10 more weeks to ensure that mature
eggs occur and the embryo’s safe development.
 With this being said, progesterone is prescribed in
IVF journey to prevent it from decreasing and
improve the preparation of the uterine lining.

27. Continued:-
 This enhances implantation success and
contributes to healthy development after
embryo transfer.
 When prescribed progesterone, start
taking it on the day of egg retrieval
unless directed otherwise by doctor.

28. Estrogen
 It is responsible for growing and maintaining
the uterine lining, particularly making it thicker
over time after fertilization so that a healthy
pregnancy can be supported.
 To support the process of IVF, estrogen
supplements prescribed so that the level of the
hormone is balanced out. The reason is that it
may change under the impact of other
reproductive hormones and procedures, so it’s
crucial to ensure the right estrogen level for
safe implantation and pregnancy development.

29. Prolactin
 Excessive prolactin affects the development of
ovarian follicles and uterine lining. While generating
milk, prolactin shuts down the woman’s ability to
reproduce. So high levels of prolactin in women who
are not pregnant or breastfeeding signal a hormonal
imbalance.
 Therefore, it’s important to track the level of prolactin
when to start IVF treatments, for which need to take
a prolactin test. It’s best to take it in the morning,
regardless of the day of cycle.
 A normal prolactin level for females before
pregnancy is <25 ng/mL. If a higher level, prescribed
medication to suppress prolactin and improve
chances of conceiving.

30. TSH (Thyroid stimulating
hormone)
 Thyroid stimulating hormone (TSH) is another
hormone that affects the reproductive system and
the woman’s ability to conceive. In particular, the
balanced levels of thyroid hormones ensure that
fertilization is successful and the woman doesn’t
experience ovarian dysfunction or polycystic ovary
syndrome, along with other reproductive issues or
problems during pregnancy.
 So taking a TSH test to determine the levels of the
hormone may be a necessity for females before
starting IVF. The test involves a blood sample taken
and can be done on any morning. The normal TSH
level for women who are not pregnant is <5.0 mIU/L.

31. hCG (Human Chorionic
Gonadotropin)
 Human chorionic gonadotrophin, also
known as hCG, is often referred to as the
pregnancy hormone. The reason is that it is
produced as the clinical pregnancy occurs
and is the highest at early pregnancy
stages. This is also the hormone that
signals pregnancy when doing a pregnancy
test.
 Along with other hormone injections, this
hormone supports the thickening of uterine
lining and the development of the embryo
after embryos are transferred

32. How long to take hormones for
IVF?
 In general, hormonal treatment for IVF starts
with suppressing natural menstrual cycle
through medication. Then, it is time to facilitate
egg production in ovaries with FSH injections
before egg retrieval. Altogether, the time of
how long to take hormones for IVF rounds to
about 2 weeks.
 Depending on what happens next, prescribed
more medication. If pregnancy takes place,
need to take progesterone to support it.
Otherwise, going for another IVF cycle, one
will have to go through the 2-week treatment
again.

33. PROTEINS
 Higher C-reactive protein levels during IVF
stimulation are associated with IVF failure.
 An inflammatory state, indicated by high CRP
levels during stimulation, may serve as a poor
prognostic indicator of IVF success.
 AOPP ( advanced oxidation protein
products) may be a potentially effective
marker for predicting the oocyte quality and
outcome of IVF, particularly in infertile women
with Endometriosis.

34. CONTINUED
 This provides a novel theoretical basis,
suggesting that anti-oxidative treatments
aimed at reducing the AOPP levels may
be a new effective strategy to promote
the maturation of oocytes and improve
the success rate of IVF.

35.  Peroxisomes proliferative-activated
receptors (PPARs) are nuclear
receptors that involved in cellular lipid
metabolism and differentiation. The
subtype γ of the PPAR family (PPARγ)
plays important roles in physiologic
functions of ovaries but not directly
related to IVF success rate.

36.  PRP (platelet rich plasma) might improve the
endometrial receptivity through the
improvement of cell proliferation,
vascularization, anti-inflammatory properties
and the reduction in the degree of fibrosis, with
the help of the concentrated peptides
(antimicrobial), GFs and cytokines in PRP.
 Matrix metalloproteinase(MMP)1, MMP3,
MMP7, and MMP26, members of MMP gene
family, involved in tissue regeneration and
wound healing via degradation of extracellular
matrix (ECM) and wound remodeling

37.  Decrease of sperm-specific
protein actin-like 7A (ACTL7A) led to
low fertilization rate, poor embryo
development, and even
infertility. ACTL7A has the potential to
be a biomarker for predicting success
rate of fertilization and effective embryo
and the possibility of embryo arrest.

38.  The combined analysis of ACTL7A,
PLC (phospholipase c ζ, and PAWP
post acrosomal sheath WW domain
binding protein) proteins in sperm
might have better prediction of IVF
outcomes.

39.  Jumonji (JARID2) protein composes a
histone methyltransferase complex called
polycomb repressive complex 2 (PRC2),
which modifies chromatin methylation to
silence many embryonic patterning genes,
acting as a negative regulator of cell
proliferation signaling. This results with a
restricted gene expression to an
appropriate cell population that is essential
for development, differentiation, and
maintenance of cell fates.

40.  Proteins on the surface of the uterine
lining cells have been identified and
some of these seem to be necessary for
normal implantation. These surface
proteins on the cells of the uterine lining
are produced by genes inside the
uterine cells.

41. THE END