Morphological study of fully and partially isolated early human follicles.
OBJECTIVE
To compare the development of fully and partially isolated human follicles by using various culture systems.


METHODS
Human ovarian material was incubated with collagenase and deoxyribonuclease. Fully and partially isolated follicles (30-50 microm) were dissected and studied under light and electron microscopy. The follicles were then cultured on and within various matrices. Fully isolated follicles were also cocultured with stromal cells.


METHODS
Rabin Medical Center, a major care and referral center.


METHODS
Women undergoing laparoscopy.


METHODS
None.


METHODS
Microscopy studies, follicular measurements.


RESULTS
Electron microscopy studies revealed an excess of lipid droplets in the granulosa cells of freshly isolated follicles. An increase in follicular size and granulosa cell number was observed only in the fully isolated follicles cultured within collagen gels for 24 hours. Most of the partially isolated follicles detached from the collagen gels. When cultured on collagen, extracellular matrix, and poly-L-lysine, both the fully and the partially isolated follicles deteriorated within the first 24 hours; coculture with stromal cells had no beneficial effect.


CONCLUSIONS
The excess in lipid droplets in granulosa cells of isolated follicles might suggest that the isolation process does not yield completely healthy follicles. However, despite this finding, our studies show that fully isolated follicles, but not partially isolated follicles, can grow within, but not on, a culture matrix.
Abir
R
R
Department of Obstetrics and Gynecology, Rabin Medical Center, Beilinson Campus and Sackler Faculty of Medicine, Tel Aviv University, Petah Tikva, Israel. ronitabir@hotmail.com
Fisch
B
B
Nitke
S
S
Okon
E
E
Raz
A
A
Ben Rafael
Z
Z
D000293:Adolescent; D000328:Adult; D002469:Cell Separation; D018920:Coculture Techniques; D003094:Collagen; D015925:Cryopreservation; D003470:Culture Media; D046508:Culture Techniques; D005109:Extracellular Matrix; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D010535:Laparoscopy; D050356:Lipid Metabolism; D008854:Microscopy, Electron; D006080:Ovarian Follicle / Q000033:anatomy & histology* / Q000502:physiology / Q000648:ultrastructure; D011107:Polylysine; D017154:Stromal Cells / Q000502:physiology
D003470:Culture Media; D011107:Polylysine; D003094:Collagen