conventional undefined mediums and unfrozen controls
(unpublished data).
In this chapter, we present a xeno-free and chemically defined
protocol for clinical-grade cryopreservation of hiPSCs along with
an undefined freezing protocol that can be used for research applications of hiPSCs.
2 Materials
2.1 Equipment
1. Class II biological safety cabinet.
2. À80
C mechanical freezer.
3. NALGENE
® Mr. Frosty freezing container or similar 1
C/
min-freezing containers.
4. Pipette aid.
5. Water bath.
6. Centrifuge.
7. Tissue culture incubator.
8. Dissecting stereomicroscope.
9. Inverted phase-contrast microscope.
10. Hemocytometer.
11. Autoclave.
12. Liquid nitrogen tank.
13. Cell storage tank.
14. Liquid nitrogen dewar.
15. Eye protection.
16. Cryogloves (face mask or goggles).
2.2 Disposables
1. Conical centrifuge tubes, 15-mL, sterile.
2. Conical centrifuge tubes, 50-mL, sterile.
3. 6- and 12-well cell culture plates.
4. Cryovials.
5. 5-, 10-, 25-, and 50-mL plastic disposable pipettes, sterile.
6. 20-, 200-, and 1000-μL pipette tips, sterile.
7. 0.2-μm sterile filter units, different membranes.
8. 0.2-μm sterile syringe filters, different membranes.
9. Kimwipes.
10. Parafilm.
11. Liquid nitrogen.
Chemically Defined Cryopreservation of hiPSCs
571
(unpublished data).
In this chapter, we present a xeno-free and chemically defined
protocol for clinical-grade cryopreservation of hiPSCs along with
an undefined freezing protocol that can be used for research applications of hiPSCs.
2 Materials
2.1 Equipment
1. Class II biological safety cabinet.
2. À80
C mechanical freezer.
3. NALGENE
® Mr. Frosty freezing container or similar 1
C/
min-freezing containers.
4. Pipette aid.
5. Water bath.
6. Centrifuge.
7. Tissue culture incubator.
8. Dissecting stereomicroscope.
9. Inverted phase-contrast microscope.
10. Hemocytometer.
11. Autoclave.
12. Liquid nitrogen tank.
13. Cell storage tank.
14. Liquid nitrogen dewar.
15. Eye protection.
16. Cryogloves (face mask or goggles).
2.2 Disposables
1. Conical centrifuge tubes, 15-mL, sterile.
2. Conical centrifuge tubes, 50-mL, sterile.
3. 6- and 12-well cell culture plates.
4. Cryovials.
5. 5-, 10-, 25-, and 50-mL plastic disposable pipettes, sterile.
6. 20-, 200-, and 1000-μL pipette tips, sterile.
7. 0.2-μm sterile filter units, different membranes.
8. 0.2-μm sterile syringe filters, different membranes.
9. Kimwipes.
10. Parafilm.
11. Liquid nitrogen.
Chemically Defined Cryopreservation of hiPSCs
571