Isolation and identification of ovarian theca‐interstitial cells and granulose cells of immature female mice

Abstract

Theca-interstitial cells (TICs) and granulosa cells (GCs) are important components of follicles that support follicle development and hormone secretion, and are considered to be important cell models for basic research. However, no method currently exists for simultaneously isolating TICs and GCs from a single ovary of the immature mouse. Here, we sought to develop such a protocol using mechanical dissection combined with brief collagenase-DNase digestion. Morphological characteristics and molecular markers were detected to identify TICs and GCs. In isolated TICs, cholesterol side chain cleavage cytochrome P450 (P450scc) was expressed abundantly, but anti-Mullerian hormone (AMH) was expressed only at very low levels. This expression profile was reversed in GCs. In addition, TICs secreted large amounts of testosterone (T) and minimal amounts of estradiol (E2 ), while the converse was found in GCs. T concentrations rose gradually in TIC culture media as the concentration of added luteinizing hormone (LH) was increased. In GCs, E2 secretion increased as the follicle-stimulating hormone (FSH) concentration increased. Thus, mechanical dissection combined with collagenase-DNase digestion is a simple, effective and reproducible method for obtaining large numbers of highly purified and hormonally stimulated TICs and GCs from one ovary.

Low Expression of Mfn2 Is Associated with Mitochondrial Damage and Apoptosis of Ovarian Tissues in the Premature Ovarian Failure Model

“…In recent years, more researchers are studying the etiologies and mechanisms of POF [ 2 , 3 , 6 ], which included the genetic contribution of the disease. Mfn2, a conserved dynamin-like GTPases that is located on the mitochondrial outer membrane and that is involved in the process of mitochondrial fusion, is conducive to the maximization of ATP production [ 27 ].…”

“…Apoptosis is an ATP-dependant process [ 36 , 37 ]. It has been shown that the apoptosis of ovarian granular cells and oocytes is the main factor in the occurrence of POF [ 2 , 3 ]. Several apoptotic pathways have been reported, including the mitochondrial, the death receptor, and the endoplasmic reticulum pathway [ 38 ].…”

“…Apoptosis is a form of cell death triggered by a variety of intracellular and extracellular signals, including mitochondrial pathways. Granulose cells (GC), a kind of functional cell, are able to synthesize many active peptides, and are involved in the synthesis of estrogen and progesterone [ 2 , 3 ]. Follicular atresia is the apoptosis of ovarian granular cells and oocytes.…”

The ovarian stroma as a new frontier

“…Of note, the concept of theca interstitial cells (TICs) has been used as a catch-all for the residual ovarian tissue husk once follicles had been punctured (Tingen et al 2011, Tian et al 2015. When cultured, theca interstitial cells from mouse ovaries take on a fibroblastlike appearance that is distinct from granulosa cells (Tian et al 2015). The heterogeneity of the TICs has been noted, with a reported shift in populations over a 12-day co-culture with follicles.…”

Human DENND1A.V2 Drives Cyp17a1 Expression and Androgen Production in Mouse Ovaries and Adrenals

“…TIC and GC were isolated from F3 wild-type and CMV-hDENND1A.V2 Tg mouse ovaries and cultured as described by Tian et al [24]. Briefly, ovaries were aseptically removed from wild-type and…”