title: 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.
authors: Department of Obstetrics and Gynecology, Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi Clinical College of Wuhan University, Enshi, Hubei, 445000, P.R. China.
authors: Yong
authors: Y
authors: Tian
authors: Wei
authors: W
authors: Shen
authors: Zhiwen
authors: Z
authors: Lai
authors: Liangyan
authors: L
authors: Shi
authors: Shuhong
authors: S
authors: Yang
authors: Ting
authors: T
authors: Ding
authors: Shixuan
authors: S
authors: Wang
authors: Aiyue
authors: A
authors: Luo
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000378:metabolism; D002469:Cell Separation; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology* / Q000378:metabolism; D007986:Luteinizing Hormone / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012741:Sexual Maturation / Q000235:genetics; D013799:Theca Cells / Q000166:cytology* / Q000378:metabolism; D059467:Transcriptome
chemical_list: D004958:Estradiol; D054304:Anti-Mullerian Hormone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone
keywords: granulosa cells; identification; isolation; steroidogenesis; theca-interstitial cells