Morphological and ultrastructural analysis of sheep primordial follicles preserved in 0.9% saline solution and TCM 199.
The objective was to determine the morphological and ultrastructural features of sheep primordial follicles preserved in either 0.9% saline solution or TCM 199 at different temperatures. Soon after death, the ovarian pair of each ewe (n = 5) was divided into 25 fragments. One fragment was immediately fixed for morphological evaluation (control). The other 24 fragments were randomly distributed in tubes containing 2 ml of 0.9% saline solution or TCM 199 and maintained at 4, 20 or 39 degrees C for 2, 4, 12, or 24h. Based on histological assessment, storage of ovarian fragments in 0.9% saline solution at 20 degrees C for up to 24h and in both solutions at 39 degrees C for 4, 12 or 24h increased (P < 0.01) the percentage of degenerate primordial follicles compared with controls. In contrast, preservation at 4 degrees C in both solutions, kept the percentage of morphologically normal primordial follicles similar to control values. Although histological integrity of primordial follicles was maintained in fragments stored at 20 degrees C for up to 24h in TCM 199, these results were not confirmed by ultrastructural analysis. Based on transmission electron microscopy, only primordial follicles stored at 4 degrees C for up to 24h, at 20 degrees C for up to 12h and at 39 degrees C for up to 2h in both solutions were ultrastructurally normal. In conclusion, sheep primordial follicles were successfully preserved at 4 degrees C for up to 24h, at 20 degrees C for up to 12h and at 39 degrees C for 2h in 0.9% saline solution or TCM 199.
Matos
M H T
MH
Laboratory of Manipulation of Oocytes Enclosed in Preantral Follicles, LAMOFOPA, Faculty of Veterinary, State University of Ceará, Av. Paranjana 1700, Campus do Itaperi, Fortaleza 60740-000, CE, Brazil. htmatos@yahoo.com
Andrade
E R
ER
Lucci
C M
CM
Báo
S N
SN
Silva
J R V
JR
Santos
R R
RR
Ferreira
M A L
MA
Costa
S H F
SH
Celestino
J J H
JJ
Figueiredo
J R
JR
D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure; D018870:Endoplasmic Reticulum, Rough / Q000648:ultrastructure; D005260:Female; D006107:Granulosa Cells / Q000648:ultrastructure; D008854:Microscopy, Electron; D008928:Mitochondria / Q000648:ultrastructure; D019188:Organ Preservation Solutions; D009994:Osmolar Concentration; D006080:Ovarian Follicle / Q000648:ultrastructure*; D012756:Sheep / Q000033:anatomy & histology*; D012965:Sodium Chloride; D014021:Tissue Preservation / Q000379:methods / Q000662:veterinary*
D019188:Organ Preservation Solutions; D012965:Sodium Chloride