Abstract


Moafaq Samir*,1, Dareen Mattar2 and Philip Gerald Knight2
1. University of Wasit, Wasit, Iraq; 2. University of Reading, Reading, United Kingdom

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Moafaq Samir*,1, Dareen Mattar2 and Philip Gerald Knight2
1. University of Wasit, Wasit, Iraq; 2. University of Reading, Reading, United Kingdom

The ovary is classified as one of the most dynamic tissues in the body because of the recurrent tissue growth and involution every month from puberty till menopause. During ovulation macrophages invade the ovary and secret pro-inflammatory cytokines such us TNF-α and IL6 that have local actions on ovarian cells1. The infiltration of macrophages into ovarian tissue as well as their presence in the female reproductive tract is strong evidence for their role in the female reproductive process2. Here we investigated (1) the presence of macrophages in bovine ovary; (2) the effect of macrophages on migration of luteinized granulosa, theca and stroma cells evaluated using an in vitro ‘wound healing’ assay.

Bovine ovaries were collected from an abattoir and the CL stages categorized by gross morphology. After cryosectioning tissue was stained with macrophages markers CD68 and MHCII. Bovine monocyte-derived macrophages were prepared from citrated blood. The macrophages were cultured with 10% FCS for seven days then treated with LPS (10µg/ml). Conditioned medium was collected and the cells were processed for RNA extraction and RT-PCR. LPS promoted a significant upregulation in expression of TNF-α, IL6, and TLR4 in macrophages (p<0.01). For wound-healing (‘scratch’) assays freshly harvested granulosa, theca and stroma cells were cultured under luteinizing conditions (medium with10% serum) for two days with/without LPS-treated macrophages or macrophage-conditioned media. A ‘scratch’ was made in the near confluent monolayer and cell migration (% wound closure) assessed over 24h using time-lapse microscopy. Immunohistochemistry showed that macrophages were present in the whole ovarian sections with the highest density evident in early stage CL. Macrophages accelerated wound healing by luteinised theca and stroma cells (30% compared with control; P<0.01), however granulosa cells showed no migratory activity regardless of macrophage presence. With stroma cells media from the LPS-treated macrophages accelerated wound healing by 20% compared with control (P<0.01) but no effect was seen with theca cells. LPS treatment alone had no effect on wound healing by either theca or stroma cells.. Results indicate a local stimulatory effect of macrophages on ovarian cell migration that may contribute to CL formation; this effect is likely mediated by pro-inflammatory cytokines TNF-α and IL6.

Reference: (1) Turner et al., Reproduction, 141, 821-31. (2) Miller and Hunt, Life Sci, 59,1-14.

Nothing to disclose MS, DM, PGK

Disclosure: MS: , Merck & Co.. Nothing to Disclose: DM, PGK

Please take note of the Endocrine Society's News Embargo Policy at: https://www.endocrine.org/news-room/endo-annual-meeting

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