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RDS Test 4 > B8.065 Assisted Reproductive Technology > Flashcards

B8.065 Assisted Reproductive Technology Flashcards

1
Q

what is ART

A

in vitro handling of both human oocytes & sperm OR embryos for purpose of establishing a pregnancy

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2
Q

types of ART

A 
IVF
embryo transfer
gamete and embryo cryopreservation
oocyte and embryo donation
gestational carriers
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3
Q

what is NOT ART

A

intrauterine insemination

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4
Q

structure and function of FSH

A

heterodimeric glycoprotein (a chain similar to other anterior pituitary hormones, B chain makes it unique)
3-4 hour half life
stimulates granulosa cells > critical regulator of follicular development

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5
Q

regulation of FSH levels

A

suppressed by rising E levels from developing follicle

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6
Q

when is FSH the highest during the cycle

A

cycle day 3
level reflective of maternal age and ovarian reserve (ie, it will be higher if you have less eggs bc there is less negative feedback)

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7
Q

how is FSH used in ART

A

recombinant injection used to stimulate oocyte development

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8
Q

function of E2 in ART

A

most important product of granulosa cell from developing follicle
given to patients to thicken endometrial lining (but can slow follicular growth)

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9
Q

when does E2 peak during the cycle

A

late follicular phase

peak level around 200

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10
Q

structure and secretion of LH

A 
heterodimeric glycoprotein (similar to HCG)
20 min half life
secreted in a pulsatile manner
-follicular q 90 min
-luteal q 2-3 hr
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11
Q

function of LH in cycle

A

stimulates mature granulosa cells

stimulates lutenized cells of corpus luteum to produce progesterone

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12
Q

use of LH in IVF

A

dont give LH bc it rapidly degrades, give hCG instead (longer half life and less expensive)
given to mature and/or release oocyte with fertility treatment (for egg retrieval)

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13
Q

what happens when the oocyte responds to the LH surge

A

resumption of oocyte meiosis
completion of metaphase I
polar body extrusion
metaphase II arrest

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14
Q

response of follicle to LH surge

A

breakdown of follicular wall
cumulus-oocyte complex ovulated
lutenization of remaining granulosa and theca cells

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15
Q

when does ovulation occur with respect to the LH surge

A

34-36 hours after

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16
Q

function of the corpus luteum

A

secretes P4 and lipid droplets accumulate in cells

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17
Q

function of P4

A

prepared endometrium for implantation

maintains fetal-placental unit until placenta takes over (8-10 wks gestation)

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18
Q

P4 in ART

A

given to patients in luteal phase of fertility treatment to support possible pregnancy

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19
Q

steps of fertilization (start w ovulation)

A
  1. oocyte remains in ampulla of fallopian tube for 3 days (waiting for sperm)
  2. sperm reaches zona pellucida
  3. acrosomal reaction
  4. sperm head binds to sperm receptor
  5. fusion of sperm and egg plasma membranes
  6. cortical reaction
  7. zona pellucida hardening
  8. oocyte nucleus completes maturation to yield female pronucleus and second polar body
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20
Q

what is the acrosomal reaction

A

leads to breakdown of zona pellucida

acrosome reacts with perivitelline space

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21
Q

what is the cortical reaction

A

microvilli on oocyte surface surround sperm head, oocyte releases cortical granules
THIS LEADS TO zona pelludica hardening which prevents polyspermia

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22
Q

prezygote

A

fertilized pronuclear oocyte with male and female pronuclei

23
Q

zygote

A

fertilized one cell stage without nuclei before cell cleavage

24
Q

preembryo

A

conceptus during cleavage stages and until 14d post-fertilization (when primitive streak develops)

25
Q

morula

A

preembry stage from 16 cell to blastocyst formation

72-96 hours post insemination

26
Q

blastocyst

A

postmorula preembryonic stage in which fluid filled blastocele develops

27
Q

embryo

A

conceptus from primitive streak until major organ development at the end of the 8th post ovulatory week

28
Q

when does hatching of the blastocyst occur

A

day 6-7

29
Q

when does implantation occur

A

day 8-9

30
Q

embryo development within the fallopian tube

A

transport area of egg/sperm
fertilization occurs in ampulla
location of all embryo development until blastocyst stage

31
Q

outcome of blastocyst components

A

inner cell mass > fetus
outer cell mass > placenta and fetal membranes
hatches from zona pellucida around day 7 and implants in uterus

32
Q

process of implantation

A
  1. requires prior conditioning by P4 (similar success rates with various protocols, just important that it is given)
    - P4 causes stromal cell swelling and accumulation of glycogen, lipids, and protein
    - hCG from blastocyst stimulates CL to secrete P4
  2. blastocyst approaches uterine wall (adplantation)
  3. blastocyst attaches to uterine wall (adhesion)
  4. invades trophoblast and embeds
33
Q

when is ART needed

A

bilateral tubal disease
severe male factor (total motile sperm < 5 mil)
genetic disorder
failed other treatment modalities

34
Q

what is IVF?

A

in vitro fertilization
eggs surgically removed from ovary
sperm mixed with egg outside of body
fertilized egg (embryo) placed into uterus
-<5% of all fertility treatments in the US involve IVF

35
Q

describe controlled ovarian hyperstimulation

A
  1. daily gonadotropin injection (mainly FSH) for 10 days to mimic follicular phase
  2. GnRH agonists given to prevent ovulation
  3. lead follicles 18-22 mm in diameter indicate possible MII oocyte
  4. hCG given for oocyte maturation
  5. retrieval 34-36 hours after hCG
36
Q

how is oocyte retrieval performed

A

transvaginal ultrasound guided
performed under conscious sedation
no betadine for vaginal prep (just normal saline + abx afterward to prevent PID)
needle attached to pump to suction out the follicular fluid and cumulus-oocyte complex

37
Q

fertilization in ART

A

ICSI or conventional

  1. activated oocyte releases 2nd polar body (1N DNA)
  2. male and female pronuclei form and approach each other
  3. 2 pronuclei in 6-10 hours suggests fertilization
    - 1 or >2 pronuclei suggests abnormal fertilization
38
Q

what is ICSI and when is it indicated

A

intracytoplasmic sperm injection
-injection of sperm into mature oocyte (at metaphase II)
indicated with severe male factor infertility (TMS < 5 mil)

39
Q

steps of ICSI

A
  1. semen processing: centrifugation to remove seminal fluid
  2. oocyte preparation: gonadotropin stimulation, hCG injection, retrieval 34-36 hrs after hCG
  3. microinjection setting: inverted microscope
  4. sperm selection: put sperm in a dense solution to stop movement
  5. oocyte maturity: inject at MII
  6. oocyte penetration
  7. evaulation of fertilization
40
Q

when do you use conventional insemination for IVF

A

no significant male infertility

no significant concern for failed fertilization

41
Q

steps of conventional insemination

A
  1. fresh semen sample collected by masturbation
  2. select highly motile sperm with normal morphology
  3. incubated in protein rich media to achieve capacitation
  4. 50-100,000 motile sperm incubated with each oocyte for 12-20 hours
42
Q

embryo culture for IVF

A

replicate conditions in fallopian tube

  • temperature
  • electrolyte, protein, carbohydrate concentrations
  • pH
  • osmolarity
  • exposure to light
43
Q

what is sequential media

A

mimics native environment

  • D1-2 nonessential amino acids and pyruvate
  • D3+ essential amino acids and glucose
44
Q

when do you do an embryo transfer

A

@ the blastocyst stage (day 5-6)

  • higher implantation rate
  • lower twin rate
  • ability to assess “true” viability
45
Q

how are embryo transfers done

A

US guided
placement 2 cm from fundus
technique associated w success rates (don’t bang around in there)
embryo selected by blastomere number/uniformity and degree of fragmentation

46
Q

factors that negatively impact IVF success rate

A 
advanced maternal age
cigarette smoking
obesity
communicating hydrosalpinges
increasing duration of infertility
low ovarian reserve
47
Q

trouble with advanced maternal age

A

cohort of growing follicles diminished
chromosomal segregation errors in female meiosis associated with embryo aneuploidy
-most errors in meiosis 1
-defects in spindle assembly

48
Q

discuss tubal infertility

A

30% of female factor infertility

salpingitis primary etiology

49
Q

diagnosis of tubal infertility

A

hysterosalpingogram gold standard

50
Q

how to work with tubal inferility issues

A

IVF higher success rate than tubal surgery
salpingectomy before embryo transfer recommended if communicating hydrosalpinx
-50% increase in success rate
-removes reflux of cytotoxic fluid from hydrosalpinx to uterine cavity

51
Q

preimplantation genetic testing for monogenic disorders

A

direct testing of IVF embryos, typically for single, inherited genetic diseases (CF)

52
Q

preimplantation genetic testing for aneuploidy

A

direct testing of IVF embryos for large-scale chromosome abnormalities (trisomy, deletions, duplications)
may improve success rates for women with advanced for maternal age and/or women with multiple number of high quality blastocysts

53
Q

techniques for preimplantation genetic testing

A

PCR

  • single gene defects
  • primers are developed to surround mutation
  • affected individuals saliva tested to ensure mutation can be detected using designed primers

RDS Test 4 (17 decks)

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