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PSIO 469 Exam 03 > Lecture 15: Gamete and Embryo Cryopreservation > Flashcards

Lecture 15: Gamete and Embryo Cryopreservation Flashcards

1
Q

Sperm can be frozen “INDEFINTELY” with cryoprotectant and stored in liquid nitrogen?
T/F

A

True

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2
Q

When was the first human SPERM cryopreservation?

When was the first baby born from frozen sperm?

A
  • 1950’s

- 1953

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3
Q

What reasons to people freeze sperm in a sperm bank?

6

A
  • Cancer-related therapy
  • Testicular or Prostate surgery
  • Vasectomy
  • High-risk occupational exposure
  • IUI or ART
  • Varied time intervals of partners
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4
Q

What is “Sperm Banking?”

A

Saving sperm for your own future use

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5
Q

What is “Sperm Donation?”

A

Allow your sperm to be used by someone else

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6
Q

What reasons do people use frozen sperm from donations?

3

A
  • Used by couples who have male infertilities
  • Couples who have genetic abnormalities
  • Single Women or Lesbians
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7
Q

What are the qualifications of donor sperm?

3

A
  • Around ages 20-39
  • Pyschological, genetic and medical screening
  • STD screening
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8
Q

Donor sperm are frozen and quarantined for ___ months to permit _______ of STD’s

A
  • 6

- re-testing

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9
Q

What are the financial compensations for sperm donation?

A
  • $125 per donation
  • Donate 2-3 times per week (24-48 abstinence period)
  • Expected Participation is 9-12 months
  • 1099 tax form
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10
Q

What is “cryopreservation?”

A

Process where cells or whole tissues are preserved by cooling to low sub-zero temps in liquid nitrogen

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11
Q

What is the temperature they keep the sperm in liquid nitrogen?

A
  • (77K)
  • (-196 C)
  • (-321 F)
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12
Q

What 2 ways are cells protected by freezing injury?

A
  • Controlled cooling (& thawing) rate

- Cryoprotectants (a form of anti-freeze)

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13
Q

What is the Slow Freezing Danger zone between?

A
  • 0 to -60 C
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14
Q

What happens between 0 to -5 C?

A

Cells and surrounding medium remain unfrozen and super-cooled

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15
Q

What happens between -5 to -15 C?

A
  • Ice forms in external medium spontaneously or by controlled seeding
  • Contents of cells remain unfrozen and super-cooled
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16
Q

What happens between -15 to -60 C?

A

Contents of cell freeze

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17
Q

What happens during Extracellular Ice Formation?

A
  • Ice crystals form in EC space
  • Cell membranes is permeable to liquid water but not ice
  • Water expands as it freezes, EC ice crystals physically compresses cells
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18
Q

Why can Extracellular Ice cause mechanical damage to cell membrane?

A

Because the ice crystals are sharp

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19
Q

What happens during Solution Effects?

A
  • Ice Crystals grow in freezing EC water

- Electrolytes and other solutes are excluded, causes them to become concentrated in reaming liquid water

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20
Q

During Solution Effects, Extracellular Fluid becomes HYPOtonic to cell.
T/F

A

False, it becomes HYPERtonic

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21
Q

What is “Osmolality?”

A

Concentration of particles dissolved in a fluid

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22
Q

What happens during Dehydration?

A
  • Water migrates out of cell due to EX ice formation coupled with increasing solute concentration in liquid water
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23
Q

What is the fate of the cell during Dehydration?

What is the fate of the cell during Cytolysis?

A
  • Cells shrinks from efflux of water

- Cell ruptures from influx of water

24
Q

What happens during Intracellular Ice Formation?

A
  • Intracellular ice is always fatal to cell, because organisms and tissues can tolerate some EC ice
25
Q

What happens during The Rate of Cooling?

A
  • Water moves out of the cells by osmosis due to increasing osmolality of EC environment as water is incorporated into ice crystals
26
Q

The SLOWER the cooling rate the ____ intracellular water has to move out

A

Longer

27
Q

What are “Cryoprotectants?”

A

Substance(s) used to protect biological tissue from freezing damage

28
Q

What Permeates when in Cryoprotectants?

3

A
  • Small molecules diffuse across membranes
  • Form hydrogens bones with water to slow or prevents both IC and EC ice crystallization
  • Protecting cells from solution effect by diluting electrolytes which are toxic to cells
29
Q

What are some electrolytes that are toxic to cells?

Hint: there are at least 3 on this list

A
  • Propylene glycol
  • Glycerol
  • DMSO
30
Q

What Non-Permeates when in Cryoprotectants?

A
  • Large molecules remain in EC
  • Slow or prevent EC ice formation
  • Can assist controlled dehydration of cells
  • Used in combination with premating molecules
31
Q

Optimal rate differs between _____ of differing _____ and ____ permeability.

A
  • cells
  • sizes
  • water
32
Q

What are the 4 steps of the Sperm Cryopreservation Standard Protocol.

A
  1. Semen analysis of fresh sample
  2. Divide entire specimen into vials containing equal number of sperm
  3. Add cyroprotectant and slowly freeze in liquid nitrogen
  4. Thaw one aliquot, remove cyroprotectant and repeat semen analysis
33
Q

Frozen is better than Fresh Sperm.

T/F

A

False. Fresh is better

34
Q

What is the post-thaw motility of humans sperm range?

A

20-65%

35
Q

What reasons are their loss of motility by post-thaw?

3

A
  • Decreased integrity of membranes
  • Cryo-damage to membranes of IC compartments
  • Altered energy metabolism and synthesis
36
Q

When was the first baby born from frozen embryo born?

What did Italy banned embryo freezing?

A
  • 1984

- 2004

37
Q

Frozen embryo transfer rate is similar to or better than fresh transfer.
T/F

A

True

38
Q

With Oocyte freezing:

1) What is the size of the sperm and head?
2) What is the size of the Blastomere?
3) What is the size of the Oocyte?

A

1) 5-60 picometers; 5x3 picometers
2) 40 picometers in diameter
3) 130 picometers in diameter

39
Q

Gas barrier forms between a hot surface (_____) and boiling liquid (______) if temperature it great enough.

A

Warm egg; liquid nitrogen

40
Q

Gas barrier greatly _____ heat transfer between the two which allows liquid to ____ _______ and consequently hot surface to remain hot longer

A

slow; last longer

41
Q

Meitotic Spindle is highly sensitive to temperature.

T/F

A

True

42
Q

Microtubules and Microfilaments damage may lead to ____ division of ______

A

Abnormal; Chromosomes

43
Q

Zona Pellucida hardening through a ______cortical granule release

A

premature

44
Q

Cortical reaction normally occurs BEFORE fertilization to prevent polyspermy.
T/F

A

False; occurs AFTER fertilization

45
Q

Why do women freeze their eggs for Egg Banks?

4

A
  • they are cancer patients
  • women who want to delay reproduction
  • failure to obtain sperm on retrieval day
  • ethical objections to embryo cryopreservation
46
Q

Egg donation involve fresh egg donation more than embryo freezing.
T/F

A

True

47
Q

What are the criteria for egg donation and compensation?

A
  • ages 20-30
  • physiological, genetic and medical screening
  • $3,500-$10,000 per donation
  • May donate 6 times in their lifetime
48
Q

What is “Vitrification?”

A

Process of converting material into glassy, amorphous solid free from crystalline structure

49
Q

What is the difference between Slow freeze and vitrification in regards to Saline solution?

A

Vitrified solution is clear

50
Q

What is “Autotransplanation?”

A

transplant tissue back into same individual

51
Q

What is “Allotransplantation?”

A

Transplant tissue into the same species but a different individuals

52
Q

What is “Xenotransplantation?”

A

Transplant tissue from one species to another

53
Q

What is “Heterotopic?”

A

Transplant tissue so it is not in its normal position

54
Q

What is “Orthotopic?”

A

Transplant tissue back into natural position

55
Q

What is “Vascular Anastomosis?”

A

Connect two cut or separate blood vessels to form a continuous channel

PSIO 469 Exam 03 (7 decks)

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