4. For aliquoting Custodiol medium: PS Multipurpose Container, 30 mL (plain label, sterile, V-bottom) – white cap
(Greiner Bio-One).
5. For aliquoting cryopreservation medium: 50 mL conical centrifuge tubes—blue caps (Vitrolife Labware).
6. Serological pipettes (sterile), 10 and 25 mL (Vitrolife
Labware).
2.2 Materials Needed
for the Cryopreservation
Procedure
1. Clinastar forceps, 11.5 cm of steel, single sterile packed.
2. Feather disposable scalpel, no. 22 (Feather), single sterile
packed.
3. PFM biopsy punch with Plunger-System, 2 mm, sterile.
4. Nunc cryo vials, 1.8 mL (Thermo Scientific Nunc), sterile.
5. Collection dishes, 90 Â 20 mm (Vitrolife Labware), sterile.
6. Custodiol and cryosolution (see above).
7. Oosafe sperm collection cup (sample container), 80 mL
(Gynemed GYOOPW-SC01), single sterile packed.
8. Surgicare gynecological powder-free gloves, single sterile
packed.
2.3 Materials Needed
for the Thawing
Procedure
1. Dulbecco’s phosphate-buffered saline (DPBS) solution, CTS
grade, sterile.
2. Human serum albumin (HSA) (FUJIFILM Irvine Scientific,
Santa Ana, CA, United States), sterile.
3. Sucrose ACS reagent grade, sterile.
4. Corning Falcon High-Clarity Polypropylene Centrifuge Tube,
Conical Bottom, 15 mL, sterile.
5. CELLSTAR 6-well plates (Greiner Bio-One), sterile.
6. Carl Roth Rotilabo
®
-disposable tweezers, single sterile packed.
7. Falcon cell strainer, 70 μm of Nylon (BD Becton, Dickinson,
and Company), single sterile packed.
8. Collection dishes, 90 Â 20 mm (Vitrolife Labware), sterile.
9. Oosafe sperm collection cup (sample container), 80 mL
(Gynemed GYOOPW-SC01), single sterile packed.
10. Acrodisc syringe filters, 0.20 μm (PALL Life Science), sterile.
11. Syringes, 20 mL, sterile.
12. Surgicare gynecological powder-free gloves, single sterile
packed.
2.4 Viability
Assessment (Calcein
Staining)
1. Nunclon delta-treated four-well IVF dish (Thermo Fisher Scientific), sterile.
2. AIM-V serum-free medium (Life Technologies, Invitrogen),
sterile.
Cryopreservation and Thawing of Human Ovarian Tissue
487
(Greiner Bio-One).
5. For aliquoting cryopreservation medium: 50 mL conical centrifuge tubes—blue caps (Vitrolife Labware).
6. Serological pipettes (sterile), 10 and 25 mL (Vitrolife
Labware).
2.2 Materials Needed
for the Cryopreservation
Procedure
1. Clinastar forceps, 11.5 cm of steel, single sterile packed.
2. Feather disposable scalpel, no. 22 (Feather), single sterile
packed.
3. PFM biopsy punch with Plunger-System, 2 mm, sterile.
4. Nunc cryo vials, 1.8 mL (Thermo Scientific Nunc), sterile.
5. Collection dishes, 90 Â 20 mm (Vitrolife Labware), sterile.
6. Custodiol and cryosolution (see above).
7. Oosafe sperm collection cup (sample container), 80 mL
(Gynemed GYOOPW-SC01), single sterile packed.
8. Surgicare gynecological powder-free gloves, single sterile
packed.
2.3 Materials Needed
for the Thawing
Procedure
1. Dulbecco’s phosphate-buffered saline (DPBS) solution, CTS
grade, sterile.
2. Human serum albumin (HSA) (FUJIFILM Irvine Scientific,
Santa Ana, CA, United States), sterile.
3. Sucrose ACS reagent grade, sterile.
4. Corning Falcon High-Clarity Polypropylene Centrifuge Tube,
Conical Bottom, 15 mL, sterile.
5. CELLSTAR 6-well plates (Greiner Bio-One), sterile.
6. Carl Roth Rotilabo
®
-disposable tweezers, single sterile packed.
7. Falcon cell strainer, 70 μm of Nylon (BD Becton, Dickinson,
and Company), single sterile packed.
8. Collection dishes, 90 Â 20 mm (Vitrolife Labware), sterile.
9. Oosafe sperm collection cup (sample container), 80 mL
(Gynemed GYOOPW-SC01), single sterile packed.
10. Acrodisc syringe filters, 0.20 μm (PALL Life Science), sterile.
11. Syringes, 20 mL, sterile.
12. Surgicare gynecological powder-free gloves, single sterile
packed.
2.4 Viability
Assessment (Calcein
Staining)
1. Nunclon delta-treated four-well IVF dish (Thermo Fisher Scientific), sterile.
2. AIM-V serum-free medium (Life Technologies, Invitrogen),
sterile.
Cryopreservation and Thawing of Human Ovarian Tissue
487