Comparison of two culture systems for the in-vitro growth and maturation of mouse preantral follicles.
The objective of our study was to compare to ability of collagen-treated membranes and bovine collagen gels to maintain murine preantral follicle growth and development in-vitro. To fulfill that objective, murine follicle and oocyte growth rates were followed for ten days in culture. Meiotic competence and the capacity to reach the two-cell stage after in-vitro maturation and fertilization, respectively, were then assessed. We used preantral follicles from 12 day-old CF-1 female mice that were isolated by enzymatic digestion from ovaries. Follicles were placed either on collagen-treated membranes or embedded in a bovine collagen matrix. The follicles were grown, changing the media and obtaining measurements every other day for ten days. Following culture, the granulosa-oocyte complexes were matured; the resultant metaphase II arrested oocytes were inseminated and cultured to the two-cell stage. The data was analyzed with significance considered for probability values of p < 0.05. We performed individual measurements on 650 follicles in seven separate experiments. Forty-eight hours after initial seeding and throughout the entire length of culture, both the follicles and oocytes grown in the collagen matrix were larger than follicles cultured on collagen-treated membranes (p < .0001). However, oocyte recovery rates were higher among follicles cultured on collagen-treated membranes (p < .01). Similar percentages of meiotically competent oocytes, fertilization and cleavage rates were observed in both groups. Our results show that mouse preantral follicles display a greater growth rate when grown embedded in a collagen gel matrix. This may be due to the maintenance of a normal three-dimensional organization of the follicle within the collagen matrix. However, this system does not enhance meiotic competency or fertilization rates in the mouse when compared to culture on collagen-treated membranes.
Loret de Mola
J R
JR
Division of Human Reproduction, Department of Obstetrics and Gynecology, Center for Research on Reproduction and Women's Health, Philadelphia, PA, USA.
Barnhart
K
K
Kopf
G S
GS
Heyner
S
S
Garside
W
W
Coutifaris
C B
CB
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D003094:Collagen; D005260:Female; D005307:Fertilization in Vitro; D006107:Granulosa Cells / Q000502:physiology; D008567:Membranes, Artificial; D051379:Mice; D008815:Mice, Inbred Strains; D009865:Oocytes / Q000254:growth & development*; D006080:Ovarian Follicle / Q000502:physiology*
D008567:Membranes, Artificial; D003094:Collagen