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ANAT 381 > Assisted Reproductive Technology > Flashcards

Assisted Reproductive Technology Flashcards

1
Q

describe male reproductive system

A

testis, sperm duct, penis

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2
Q

describe female reproductive system

A

ovary, fallopian tubes, uterus, vagina

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3
Q

what needs to be functional to conceive naturally

A

male and female reproductive systems - all components to conceive naturally

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4
Q

what is ART

A

Specialized treatments or methods designed to increase number of oocytes and or sperm and improve likelihood of pregnancy = assisted reproductive technologies

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5
Q

is ART just ivf

A

NAHHH
many kinds of treatments

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6
Q

when and where to use ART - infertility

A

designed initially for infertiltiy

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7
Q

when and where to use ART - cancer

A

fertility preservation
adult = get cancer and have to do chemo = toxin so damages likelihood of biological kids, freeze eggs or sperm for future use
children?=complicated because how young is too young

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8
Q

when and where to use ART - social aspects

A

preserving fertility to circumvent maternal age effect = freeze eggs while young and then have children at older age
same sex couple

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9
Q

how do same sex couples have children

A

fem/fem = get sperm donor - easier
male/male = more complicated - need egg bank and surrogate

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10
Q

when and where to use ART - prevention

A

prevention of inherited diseases and genetic disorder by preforming pgt - test embryos

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11
Q

define term infertile

A

couples who have not been able to conceive after one year (12 months) of natural unprotected intercourse

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12
Q

what is subfertile

A

few couples are completely unable to have children - many are considered to be subfertile = have reduced ability to conceive and produce children = not completely infertile, can conceive with help

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13
Q

Clinical definition of infertility according to WHO

A

disease (no fault) of the reproductive system define by failure to achieve a clinical pregnancy after 12 months of more of regular unprotected sexual intercourse

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14
Q

what is normal fertility - parameters

A

50% after 3 months
70-80% within first 6 months
85% conceive by 12 months
92% after 2 years

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15
Q

describe likelihood of fertility - percentage

A

likelihood of fertility decreases to 1% after 3 years of intercourse without contraception
becomes more difficult, should approach fertility doc after 1 year

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16
Q

describe fertile interval

A

6 days prior to ovulation to day of ovulation

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17
Q

when is highest probability of conception

A

intercourse 1-2 days prior to ovulation

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18
Q

what is normal fertility dependent on

A

maternal age mostly, lesser extent = paternal age

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19
Q

describe life of sperm/ovum

A

sperm lives 3-5 days in fem repro tract
ovum survives 12-24 hrs after ovulation
cannot figure out exact implantation window but around ovulation = best

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19
Q

etiology of fertility

A

before = used to be considered only the females fault, but now know men can contribute
male factor = 30%
female factor = 30%
combined = 10%
unexplained = 25%
other =5%
must check males and females at clinic

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20
Q

what does detection of certain genetic causes of male infertility allow for

A

to be informed about potential to transmit genetic abnormalities that may affect health of offspring

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20
Q

describe male infertility - conditions (categories)

A

some conditions = identifiable (can see problem) and reversible (like surgery)
Other conditions = identifiable but not reversible

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20
Q

describe summary of investigations in the man - fertility

A

anatomical investigation = if abnormal = surgery
semen analysis = if positive antibody reaction - treat, if infection = treat, if low count = improve lifestyle and can send to art
hormone analysis = if abnormal = treat if can
if no sperm = get donation
CAN ALL BE SENT TO ART

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20
Q

what is male infertility due to

A

deficiencies in sperm
production
blockage of the sperm delivery system
antibodies against sperm
injury to testicle
disorders of hormone production
poor descent of one or both testes
the presence of a varicocele

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21
Q

name the most commonly identifiable female factors for infertility

A

ovulatory disorders - 25% -do not ovulate normally
Endometriosis –15%
Pelvic adhesions –12%
Tubal blockage –11%
Other tubal abnormalities –11%
Hyperprolactinemia (high levels of prolactin in the blood)– 7%

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22
Q

describe summary of investigations in the woman - fertility

A

swabs and cervical smear = if abnormal treat- fix before ART
fallopian tubes = if abnormal = surgery then art
ultrasound = if abnormal surgery
Hormone levels = if abnormal = surgery or medical treatment, if no eggs = egg donations
CAN ALL BE SENT TO ART

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23
Q

infertility in population

A

1 in 6 couples will face a fertility issue in their reproductive period

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24
Q

What are the options for Infertile couples

A

adopt
no children
ART

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25
Q

who may benefit from ivf - 5

A

Blocked or damaged fallopian tubes or no fallopian tubes
male factor infertility = low sperm count or poor motility
women with ovulation disorders , premature ovarian failure, uterine fibroids
Individuals with genetic disorders
sometimes unexplained fertility

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26
Q

describe ART - where is it now - study prediction

A

nearly 170 to 400 mil people in 2100 (3% of world) may be alive as result of ART

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27
Q

describe ART - where is it now - to date

A

MORE THAN 5 mil ivf babies born

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28
Q

describe ART - where is it now - globally

A

ivf available in almost all parts of globe
doesnt mean its accessible tho - like someplaces = free while others not free = very expensive everywhere

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29
Q

describe normal egg development

A

females release one egg per month, sometimes 2 oocytes = frat twins
need one good egg (and one good sperm = pregnancy)

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30
Q

what is involved in ivf procedure - gen - 5 “points”

A

monitor and stimulate development of healthy eggs in ovaries - want as many as possible
collect eggs
secure sperm
combine eggs and sperm together in lab and provide appropriate environment for fertilization and early embryo growth (culture)
transfer embryos back into uterus

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31
Q

what is step 1 of IVF procedure - describe all

A

Fertility med prescribed to stimulate ovary - egg production
multiple eggs desired since some eggs will not develop or fertilize after retrieval
transvaginal ultrasound used to examine ovaries
blood tests = check hormone levels

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32
Q

describe afc - step1 ivf

A

count amount of follicles = inside has egg

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33
Q

describe standard ivf protocol - step 1

A

clinically prescribe to patients
oral contraceptive= administered on day 1 or 2 of menstrual cycle to prevent formation of cysts and schedule time of treatment - continued for 10-21 days - 3 weeks
designed per person tho - grow follicles and mature eggs - recruitment of follicles

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34
Q

how many injections - ivf step 1

A

10 days of injections
Subcutaneous
Financially, emotionally and physically hard

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35
Q

describe ultrasound image of follicular dev - step 1 ivf

A

look at size of follicles and measure
18-20ml = time to extract egg
follow with ultrasound

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36
Q

what is step 2 of IVF procedure - describe all

A

eggs retrieved through minor surgical procedure that uses ultrasound imaging to guide a hollow needle through pelvic cavity to remove eggs
medication provided to reduce and remove potential discomfort

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37
Q

describe aspects of oocyte retrieval - step 2 ivf

A

transvaginal ultrasound guided retrieval procedure
probe needle = bit of suction. to aspirate eggs and fluids
aspirate growing follicles one by one - hopefully good egg growing

38
Q

what is step 3 of IVF procedure - describe all

A

male asked to produce sample of sperm - prepared for combining with eggs

39
Q

what is step 4 of IVF procedure - describe all

A

insemination = sperm and eggs mixed together and stored in lab to encourage fertilization
some cases where there is lower probability of fertilization due to male factor (low sperm count) = icsi used = single sperm injected directly into egg
eggs are monitored to confirm fertilization and cell division are taking place
once fertilization = now considered embryos

40
Q

what is step 5 of IVF procedure - describe all

A

embryos transferred into womans uterus 3-5 days after egg retrieval and fertilization
catheter or small tube inserted into uterus to transfer embryos
procedure painless for most women - some may experience mild cramping
if procedure successful = implantation occurs around 6-10 days following egg retrieval

41
Q

step 5 of IVF procedure - describe culture gen

A

culture in vitro- can transfer embryo at any stage but usually wait till blastocyst, and must choose best embryo

42
Q

describe lab aspects - oocyte retrieval and identification

A

extract eggs and pass down to embryologist in lab
look at it under microscope = find eggs and see if good

43
Q

how to score oocytes - lab aspects

A

cumulus spread = mature
disgard morphologically abnormal oocytes = degenerated, pathenogenesis, big polar bodies

Germinal vessicle oocyte = immature
m1 oocyte = no gv, no pb
m2 oocyte = first pb- mature
weed out abnormal sizes/shapes = only take normal ones
Nuclear and cytoplasmic maturation

44
Q

describe sperm analysis - gen

A

once have good egg then do sperm analysis
semen assessed for many factors
look and then score and decide which method to use for fertilization

45
Q

describe sperm analysis - what is it assessed for - ART

A

count=total number of sperm
motility = % of total viable sperm
Progression = speed of forward movement
abnormality= % of abnormal sperm

46
Q

describe sperm analysis - WHO parameters

A

sperm conc = 15mil/ml or more (usually 3-5ml)
total motility = 40% or more
progressive motility = 32% or more
morphology = 4% or more normal forms (strict criteria)

47
Q

describe sperm preparation

A

wash sperm, swim up technique
percoll gradient
sucrose gradient
pure sperm
or just buy protocol

48
Q

describe sperm morphology

A

All Sperm are Normal
but with Different Sizes
(the length of a healthy sperm tail can be from 40
to 250 μm)
abnoormal= big head, small head, dull head, 2 tails, pointy head
should be moving with tail forward and fast

49
Q

what is globozoospermia

A

round headed sperm with no acrosome
cannot fertilize egg
Chromosomal abnormal = tetraploid, so embryo produced would be genetically abnormal

50
Q

describe insemination

A

spem samples provided on day of egg collection = prepared in labelled tubes to separate out live motile sperm
each egg cumulus complex inseminated with 25,00- (100,000 sperm/ml) prepared sperm after 3-4 hr pre incubation of oocytes
dishes containing egg and sperm are incubated overnight in incubator - see if fertilized

51
Q

what is icsi - describe

A

insertion of a single sperm into the oocyte
bypassing all the oocyte coat penetration and gamete fusion steps (characteristic of natural fertilization)
sperms must be immobilized = nick tail
polar body must be at 6 o clock - so won’t disturb chroms

52
Q

which step is bypassed in icsi

A

sperm fusion step
male pronuclear development generally required oocyte activation but in humans

53
Q

in icsi - how is human oocyte activated

A

vigorous suction of ooplasm and sperm nuclear insertion - injection process itself activates egg

54
Q

when is icsi primarily used

A

Treatment of sever cases of male factor infertility with an abnormal semen analysis
do not need many good sperm to fertilize egg

55
Q

describe normal and abnormal fertilization -ivf

A

all morphological
presence of 2 pronuclei = normal and 2 polar bodies
zygotes with triple or more pronuclei = discarded
zygotes with normal fertilization at time of scoring transferred into new dishes containing pre incubated embryo culture medium for further developmental culture

56
Q

describe embryo culture - culture media

A

media and components crucial parts of what is needed at each developmental stages of embryos
was hard to determine bc started with mouse - but humans more complicated

57
Q

describe embryo culture - environment

A

many factors
homeostatic environment with as little stress as possible
very delicate
need to get environment right = temp and stress free

58
Q

what are incubators

A

device used to grow and maintain cell and embryo cultures
maintains optimal etmp, humidity and other conditions such as co2 and o2 content of atmosphere inside
must get it right bc ph of media is important

59
Q

describe regular air vs incubator settings

A

regular air = 78% nitrogen gas (N2) and 22% oxygen (O2), CO2 content of air is less than one tenth of 1%
incubator = with 6% CO2 and 5% O2 control - interacts with culture media used to determine the ph within system

60
Q

describe ivf incubators - gen

A

early days = glass cylinder, can open and put embryo in media then close it, stick in 37 degree ovens
now = incubators connected to computer system - if something goes wrong like concentration of gases computer will know
diff types = tri gas incubators - can open only one - so then not affect others, exposing to room temp = bad all the time

61
Q

describe scoring day 1 zygote

A

2 pronuclei
2 polar bodies

62
Q

describe scoring day 2 cleavage

A

Should get 2-4 cells

63
Q

describe scoring cleavage stage grading - gen

A

grading based on how looks under microscope
worse grade embryos probably will not develop same as a better grade embryo

64
Q

describe scoring cleavage stage grading - GRADE 1

A

Evenly sized blastomeres
spherical
Moderate refractility (not very dark)

65
Q

describe scoring cleavage stage grading - GRADE 2

A

Uneven or irregularly
shaped blastomeres
Variation in refractility
< 10% fragmentation

66
Q

describe scoring cleavage stage grading - GRADE 3

A

< 50% fragmentation
Remaining blastomeres should be in Grade 2 condition

67
Q

describe scoring cleavage stage grading - GRADE 4

A

> 50% fragmentation
 Remaining blastomeres
should appear viable

68
Q

describe scoring - day 3 cleavage stage embryo

A

Transition from maternal to zygotic gene activation at around 4-8 cells
many more cells - seems to compact

69
Q

describe scoring day 4 - compaction

A

morula
tight junctions (outer)
gap junctions (inner)
start of differentiation
tries to form blastocyst cavity, nicely compacted

70
Q

describe scoring day 5/6

A

should see icm (becomes embryo) and te (placenta)
do not develop at same rate
Choosing morphologically good quality blastocyst for transfer

71
Q

describe grading blastocyst - all

A

ICM = A- numerous and tightly packed B- several and loosely packed cells C- few cells
TE = A- many tightly packed cells organized into epithelium B- several cells organized into loose epithelium C- few cells
GRADE A - MORE LIKELY TO IMPLANT AND SUCCEED

72
Q

do all morphologically good looking embryos work well and produce good babies

A

nooooo
do not have equal potential for implantation
since just looking at morphology
do not know genetics - chromosome abnormalities

73
Q

describe real time monitoring device for embryo selection - gen

A

idea to apply time-lapse techniques in mammalian embryology is to observe some changes in structure elements or to analyze mechanism of dynamic processes including
expansion and hatching

74
Q

describe real time monitoring device for embryo selection - purpose

A

measure the length of cell cycle, compaction, blastocoel formation, and compare the effects of various culture conditions
if does a,b,c,d = best - at certain times, how long it took untill 1st cell division, how many cell divides at what time, at what rate, when blastocele cavity forms

75
Q

describe real time monitoring device for embryo selection - mechanism

A

may help to understand the mechanism of polar body
extrusion and pronuclear formation and provide a direct
evidence about the reversible nature of fragmentation

76
Q

describe real time monitoring device for embryo selection - correlations

A

early disappearance of pronuclei and onset of the first cleavage were found to be correlated with a higher number of blastomeres on day 2 after oocyte retrieval providing the predictive potential of early-stage development of human
embryos

77
Q

describe embryoscope

A

Chamber and put dish and monitor embryos = video
time lapse incubator
embryo development not uniform = different rates

78
Q

describe human blastocyst hatching

A

proteases digest proteins and zona, once blastocyst expanded= egg must hatch out for implantation
if no hatching = embyro dies

79
Q

describe laser assisted hatching

A

makes hole and wait for it to hatch out
before = used to be solution used to make hole

80
Q

name the indications of assisted hatching - 9

A

2 previous implantation failures
37 years old and over
Day 3 FSH ≥ 15 mIU/ml
Thickened ZP (≥ 15 μm thick)
< 5 cells on Day 3
> 20% of the perivitelline space occupied by extracellular fragments
IVM embryos
Thawed oocytes/embryos
Patient request

81
Q

what is ivm

A

in vitro maturation of oocytes

82
Q

why ivm - all

A

some women v sensitive to stimulation with exogenous gonadotropins
increased risk of developing ovarian hyperstimulation syndrome (ohss) = nausea, vomiting, liquid retention, shortness of breath- during ivf process
long term side effects of repeated ovarian stimulation may increase risk of ovarian, endometrial and breast cancers - some patients worry, studies have not shown this tho

83
Q

describe ivm

A

do no give hormones
immature oocytes can be matured in vitro - after releasing from antral follicles in vitro - extract cells then mature in vitro - hard
culture conditions affect oocyte maturation rate and quality during ivm - must mimic natural envir
cumulus cells associated with oocyte quality during ivm
oocyte maturation rates in vitro are different from diff reports (50-75%, immature eggs that mature, culture for 24-36 hrs, some keep till 48, but after 36 hrs probably wont mature)

84
Q

describe ivm - procedure

A

immature oocytes –> ivm –> fertilization –> culture in vitro –> cryopreseve or insert to uterus

85
Q

what is cumulative morphological assessment

A

optimal evaluation of an embryo’s potential would necessitate multiple assessments of the embryo at each of the developmental checkpoints;
Embryo development makes its transition from a single-celled zygote after fertilization of the oocyte to a blastocyst and from maternal to embryonic genome activation, initiation of protein synthesis, and cell differentiation;

86
Q

what is most predictive information regarding embryo quality today

A

conducting noninvasive evaluation of the preimplantation period in a systematic, cumulative fashion

87
Q

name all noninvasive and invasive methods to select best quality embryo - 6

A

Metabolic Parameters for assessing Embryo Viability(all non invasive)
Oxygen Consumption as an indicator of Embryo Viability
Amino Acid Turnover as a Biomarker of Embryo Viability
Gene Expression Changes during Embryo Development: New Application for Embryo Selection (will tell you which embryo best for implantation)
Real-Time Embryo Monitoring Device for Embryo Selection
Embryos biopsy and Preimplantation genetic testing for chromosome compliments

88
Q

describe embryo transfer

A

load embryos on catheter under microscope - the best looking ones
ultrasound guided embryo transfer - transvaginal = outer sheath of catheter and deposit into endometrial cavity

89
Q

what to do with rest of embryos - 2 methods

A

1 = slow freezing = slowly reduce temp - takes around 3 hrs, then stick into liquid nitro
2 = vitrification or fast freezing = more effective and better results

90
Q

describe embryo vitrification

A

shrink at blastocyst stage - liquid can cause ice crystals - when shrink = remove liquid and only have cells, put on media - buffer or washing media, equilibration solution (3-5mins, 7.5%dmso + 7.5% eg) then vitrification solution (30 sec, 0.5M sucrose, 15%dmso +15%eg) then put on freezing straw then put into liquid nitrogen

91
Q

describe storage of spare of supernumerary embryos

A

cryo
be careful when labelling - for each patient

92
Q

descrive embryo thawing

A

run through process =
tm (1 min, 1.0M sucrose) then d1 (2min, 0.5M sucrose) then d2 (3 mins, 0.25M sucrose) then wash (5 mins, buffer or washing media) then repeat wash
expands again - takes a couple of hours for embryo to be back to normal

93
Q

describe pregnancy test

A

beta hcg test at around 14 days after embryo test
if at certain number = positive for pregnancy

94
Q

descrive viability test

A

see if fetal heart beat - embryo viable then send to obstetrician

95
Q

describe historical view of human ivf

A

took a while for scientists to culture embryo in vitro
have been taken from the mother before ovulation, fertilized in vitro and grow in vitro to the eight or sixteen-celled stage in various media
discover = 1970

96
Q

describe worlds first ivf baby

A

1978
in UK - cambridge
louise brown
needed a clinician and a scientist = Patrick steptoe and robert edwards
all IVF BABIES ARE NOT INFERTILE dude

ANAT 381 (24 decks)

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