Cryopreservation of isolated ovine primordial follicles with propylene glycol and glycerol.
OBJECTIVE
To verify the viability of isolated primordial follicles to different propylene glycol (PROH) and glycerol (GLY) concentrations before and after cryopreservation.


METHODS
Isolated primordial follicles were stained with trypan blue to evaluate the effect of different PROH and GLY concentrations before and after cryopreservation.


METHODS
Laboratorio Renzo Giuliani, University of Florence, Italy.


METHODS
Thirty- to forty-day-old lambs.


METHODS
: Isolation of primordial follicles with subsequent exposure to cryoprotectant and freezing.


METHODS
Histologic structure and follicular mortality.


RESULTS
After the isolation procedure (control), the mean number of live primordial follicles/mL was 2,688 and 4,452 in the GLY and PROH groups, respectively. When GLY was used, the number of live follicles before cryopreservation was 820, 756, 640, 524, 564, and 460 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 12, 36, 100, 84, and 68 follicles/mL, respectively, with the same concentrations. When PROH was used, the number of live follicles before cryopreservation was 4,216, 3,880, 3,560, 1,812, 704, and 568 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 116, 336, 472, 360, and 244 follicles/mL, respectively, with the same concentrations.


CONCLUSIONS
Both cryoprotectants were shown to preserve isolated primordial follicles after cryopreservation.
Amorim
Christiani Andrade
CA
University of Santa Maria, Santa Maria, RS, Brazil. amorimc_kleinj@yahoo.com
Rondina
Davide
D
Rodrigues
Ana Paula Ribeiro
AP
Gonçalves
Paulo Bayard Dias
PB
de Figueiredo
José Ricardo
JR
Giorgetti
Alessandro
A
D000818:Animals; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005990:Glycerol / Q000008:administration & dosage*; D006080:Ovarian Follicle / Q000033:anatomy & histology / Q000187:drug effects* / Q000502:physiology; D019946:Propylene Glycol / Q000008:administration & dosage*; D012756:Sheep; D014022:Tissue Survival / Q000187:drug effects; D018675:Toxicity Tests
D003451:Cryoprotective Agents; D019946:Propylene Glycol; D005990:Glycerol