Effects of follicle-stimulating hormone and serum substitution on the in-vitro growth of human ovarian follicles

Abstract

In-vitro maturation (IVM) of human ovarian follicles and oocytes could benefit infertile women, and allow the development of in-vitro systems for the study of human follicular development. Little is known about the initiation of growth of primordial follicles and the regulation of early folliculogenesis. An ovarian tissue-slice culture system was used to examine the effects of media composition, follicle stimulating hormone (FSH) and serum substitution on the development of small human follicles in vitro. Human ovarian cortex biopsies were cut into small pieces and cultured for 5, 10 or 15 days. Control (non-cultured) and cultured tissue was fixed, serially sectioned, and stained. The follicles contained within the tissue pieces were counted, measured, and assessed for stage of development and viability. Comparison of the ability of alpha-minimum essential medium (alpha-MEM), Waymouth's, or Earle's balanced salt solution (EBSS) culture media (all with 10% human serum) to support follicle growth demonstrated significantly increased initiation and growth of follicles in alpha-MEM during the first 10 days of culture. The supplementation of alpha-MEM with 300 mIU/ml FSH significantly reduced levels of atresia and increased the mean diameter of healthy follicles. Follicles in tissue cultured for 10 days with human serum albumin and ITS (insulin/transferrin/selenium mix) were significantly larger, more developed and showed significantly less atresia than those cultured with serum alone. Primordial to small preantral follicles can be grown under serum-substituted conditions in tissue-slice culture, and are responsive to FSH, which is thought to be acting mainly as a survival factor at these early stages.

In Vitro Effect of Cyclic Adenosine 3′, 5′-Monophosphate (cAMP) on Early Human Ovarian Follicles

“…The effects of cAMP on follicle development in the present study were in accordance with those obtained by using FSH supplementation (2). FSH supplementation did not increase the diameters of the follicles or promote the developmental stages during the first 5 days of culture.…”

“…It would be interesting to test combined effect of cAMP and FSH on the early human ovarian follicles. FSH is involved in early folliclulogenesis in human (2). In human, 50% of follicles at the primary stage express FSH receptor (23).…”

“…Human primordial and primary ovarian follicles have been successfully cultured to secondary and occasionally to antral stages within slices of ovarian cortical tissue (2)(3)(4)(5)(6). In these cultures, follicle stimulating hormone (FSH) (2), insulin, insulin-like growth factors I and II (4), and growth differentiation factor-9 (6) have had positive effects on the growth and survival of the follicles.…”

In vitro development of primordial follicles after long-term culture of goat ovarian tissue

“…In contrast, the other treatments (FSH-7d and FSH + FGF-2) were effective in maintaining follicle ultrastructural integrity up to day 7. Our light histological results confirm earlier studies in which FSH maintained preantral follicle viability after short (Matos et al, 2007a andMatos et al, 2007c) or long-term culture (Cortvrindt et al, 1997, Wright et al, 1999and Gutierrez et al, 2000. It is suggested that the absence of FSH in the medium could compromise the diffusion of several essential chemical and physical factors through the basement membrane (Hsueh et al, 1994).…”

“…As there are FSH receptors in the granulosa cells of early follicles, it is possible that FSH may promote follicular growth and development (Méduri et al, 2002). Previous observations have showed that FSH promotes an increase in both oocyte (Itoh et al, 2002) and follicular diameter (Wright et al, 1999) after long-term culture.…”