Alginate encapsulation supports the growth and differentiation of human primordial follicles within ovarian cortical tissue.
OBJECTIVE
In vitro follicle growth (IVFG) is an investigational fertility preservation technique in which immature follicles are grown in culture to produce mature eggs that can ultimately be fertilized. Although progress has been made in growing primate primary and secondary follicles in vitro, it has been a relatively greater challenge to isolate and culture primordial follicles. The purpose of this study was to develop methods to grow human primordial follicles in vitro using alginate hydrogels.


METHODS
We obtained human ovarian tissue for research purposes through the National Physicians Cooperative from nationwide sites and used it to test two methods for culturing primordial follicles. First, primordial follicles were isolated from the ovarian cortex and encapsulated in alginate hydrogels. Second, 1 mm × 1 mm pieces of 500 μm-thick human ovarian cortex containing primordial follicles were encapsulated in alginate hydrogels, and survival and follicle development within the tissue was assessed for up to 6 weeks.


RESULTS
We found that human ovarian tissue could be kept at 4 °C for up to 24 h while still maintaining follicle viability. Primordial follicles isolated from ovarian tissue did not survive culture. However, encapsulation and culture of ovarian cortical pieces supported the survival, differentiation, and growth of primordial and primary follicles. Within several weeks of culture, many of the ovarian tissue pieces had formed a defined surface epithelium and contained growing preantral and antral follicles.


CONCLUSIONS
The early stages of in vitro human follicle development require the support of the native ovarian cortex.
Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, 303 E. Superior Street, Lurie 10-121, Chicago, IL, 60611, USA.
Monica M
MM
Laronda
Francesca E
FE
Duncan
Jessica E
JE
Hornick
Min
M
Xu
Jennifer E
JE
Pahnke
Kelly A
KA
Whelan
Lonnie D
LD
Shea
Teresa K
TK
Woodruff
D000293:Adolescent; D000328:Adult; D000464:Alginates / Q000494:pharmacology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D020100:Hydrogels; D006080:Ovarian Follicle / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010053:Ovary / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D046509:Tissue Culture Techniques; D055815:Young Adult
D000464:Alginates; D020100:Hydrogels