Attempted application of bioengineered/biosynthetic supporting matrices with phosphatidylinositol-trisphosphate-enhancing substances to organ culture of human primordial follicles.
OBJECTIVE
To improve human primordial follicle culture.


METHODS
Thin or thick ovarian slices were cultured on alginate scaffolds or in PEG-fibrinogen hydrogels with or without bpV (pic), which prevents the conversion of phosphatidylinositol-trisphosphate (PIP3) to phosphatidylinositol-bisphosphate (PIP2) or 740Y-P which converts PIP2 to PIP3. Follicular growth was evaluated by follicular counts, Ki67 immunohistochemistry, and 17β-estradiol (E2) levels.


RESULTS
BpV (pic) had a destructive effect on cultured follicles. Thawed-uncultured samples had more primordial follicles than samples cultured in basic medium and fewer developing follicles than samples cultured in PEG-fibrinogen hydrogels with 740Y-P. There were more atretic follicles in samples cultured on alginate scaffolds than in PEG-fibrinogen hydrogels, and in samples cultured in PEG-fibrinogen hydrogels with 740Y-P than in PEG-fibrinogen hydrogels with basic medium. Ki67 staining was higher in PEG-fibrinogen hydrogels than on alginate scaffolds. E2 levels were higher in thick than in thin slices.


CONCLUSIONS
PEG-fibrinogen hydrogels appear to have an advantage over alginate scaffolds for culturing human primordial follicles. Folliculogenesis is not increased in the presence of substances that enhance PIP3 production or with thin rather than thick sectioning.
Infertility and IVF Unit, Beilinson Hospital for Women, Rabin Medical Center, Petach Tikva, 49100, Israel.
Galit
G
Lerer-Serfaty
Nivin
N
Samara
Benjamin
B
Fisch
Michal
M
Shachar
Olga
O
Kossover
Dror
D
Seliktar
Avi
A
Ben-Haroush
Ronit
R
Abir
D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D015925:Cryopreservation; D005260:Female; D006801:Humans; D020100:Hydrogels; D009369:Neoplasms / Q000473:pathology; D009924:Organ Culture Techniques; D006080:Ovarian Follicle / Q000166:cytology* / Q000601:surgery; D051059:PTEN Phosphohydrolase / Q000037:antagonists & inhibitors; D010716:Phosphatidylinositols / Q000737:chemistry; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D017968:Vanadium Compounds; D055815:Young Adult
D020100:Hydrogels; D010716:Phosphatidylinositols; D017968:Vanadium Compounds; C472777:bisperoxovanadium; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human