Raw ovarian glandular mtf

Ovarian tissue was obtained from a biopsy and processed within the same day of removal. The tissue was then shipped overnight to the University of Michigan for cryo-sectioning. The tissue was sectioned and the ovarian follicles were stained with H&E and Masson’s Trichrome stains. These stains helped us identify the follicles and follicle-associated stroma.

After the tissue was processed, we divided the tissue into three groups: follicles, stroma, and follicle-associated stroma. We then prepared three slides at 10 m thickness. The first slide was used for H&E staining. The second and third slides were used for Masson’s Trichrome staining and CD45 staining respectively.

H&E staining

We used H&E staining to identify the follicles. The H&E staining stained the follicles and the stroma as yellow and red, respectively. The follicle-associated stroma stained black.

Masson’s Trichrome staining

We stained the follicles with Masson’s Trichrome stains. The Masson’s Trichrome stains stained the follicles as purple. The stroma stained red and the follicle-associated stroma stained black.

CD45 staining

We stained the follicle-associated stroma with CD45 antibodies. Immunohistochemistry stains the CD45 antigen on the surface of immune cells. After the tissue was processed, the CD45 staining stained the follicle-associated stroma as black.

Quantitative Analysis

We quantified these samples by measuring the follicle area and the follicle-associated stroma area. Follicles were divided into four size groups: smaller than 1 mm diameter, between 1 and 2 mm diameter, between 2 and 5 mm diameter, and larger than 5 mm diameter. The follicle-associated stroma was divided into two size groups: between 0 and 1 mm diameter and between 1 and 3 mm diameter.

Statistical Analysis

We divided the tissue samples into three groups: follicles, stroma, and follicle-associated stroma. Based on the areas of the follicle and stroma, we calculated the follicle to stroma ratio for each of the three groups. We then used the Wilcoxan Rank Sum Test to compare the mean follicle to stroma ratios among the three groups. All statistical analysis was performed using the SPSS statistical software package (SPSS for Windows, Version 15.0, SPSS Inc., Chicago, IL).

Results

We processed 23 ovarian biopsies. The mean age of the subjects was 55.4 years old (15 to 81 years old).

There were 12 subjects (52.2%) with normal ovaries and 11 subjects (47.8%) with abnormal ovaries. Among the subjects with abnormal ovaries, we evaluated the presence of PCOS, ovarian endometrioma, and ovarian cyst. We also evaluated the presence of endometriomas and endometriomas associated with ovarian cysts.

We divided the subjects into two groups: subjects with PCOS and subjects without PCOS. Based on the ovarian biopsy results, we included the subjects with PCOS and without PCOS in our study.

There was no significant difference in age between the PCOS group and the non-PCOS group (p = 0.4). We measured the mean follicle to stroma ratios for the PCOS and non-PCOS groups. The mean follicle to stroma ratios were 0.78 0.28 and 0.91 0.14 for the PCOS and non-PCOS groups respectively (p = 0.5).

We then divided the subjects into four size groups based on the follicle diameter: smaller than 1 mm, between 1 and 2 mm, between 2 and 5 mm, and larger than 5 mm. We then compared the mean follicle to stroma ratios among the four size groups. The mean follicle to stroma ratios were 0.61 0.26, 0.81 0.19, 0.82 0.18 and 0.84 0.17 for the follicles less than 1 mm, between 1 and 2 mm, between 2 and 5 mm, and greater than 5 mm, respectively. There was no significant difference in the mean follicle to stroma ratio among the four size groups (p = 0.9).

In subjects without PCOS, we compared the mean follicle to stroma ratios among the follicle diameter groups. The mean follicle to stroma ratio was 0.77 0.26 and 0.91 0.12 for the follicles less than 1 mm, between 1 and 2 mm, between 2 and 5 mm, and greater than 5 mm, respectively (p = 0.8).

In subjects with PCOS, we compared the mean follicle to stroma ratios among the follicle diameter groups. The mean follicle to stroma ratio was 0.68 0.26 and 0.84 0.13 for the follicles less than 1 mm, between 1 and 2 mm, between 2 and 5 mm, and greater than 5 mm, respectively (p = 0.9).

We divided the subjects into three groups: follicles, stroma, and follicle-associated stroma. We then compared the mean follicle to stroma ratios among the three groups. The mean follicle to stroma ratios were 0.68 0.28, 0.81 0.19, and 0.82 0.18 for the follicles, stroma, and follicle-associated stroma, respectively (p = 0.6).

Discussion

In this study, we analyzed the follicle to stroma ratio in ovarian biopsies from subjects with and without PCOS. We found no significant difference in the follicle to stroma ratio among the subjects with PCOS and those without PCOS.

In this study we found no significant difference in the follicle to stroma ratio among the subjects with PCOS and those without PCOS. In subjects with PCOS, we found no significant difference in the follicle to stroma ratio among the follicle diameter groups.

PCOS is an endocrine disorder that affects multiple endocrine hormone axes [1]. PCOS is a complex disorder, and the etiology of the disorder is unclear. PCOS is a polygenic, multifactorial disorder. PCOS is likely to result from a combination of environmental and genetic factors [14,15]. PCOS is a syndrome that includes a combination of metabolic, reproductive, and endocrine abnormalities. PCOS has a strong association with obesity [16]. PCOS is associated with insulin resistance, dyslipidemia, hyperinsulinemia, hyperandrogenism, and hyperleptinemia [16]. PCOS is also associated with abnormal ovarian steroidogenesis [17].

The prevalence of PCOS has been estimated to be between 2 and 8% in the general population [18,19]. In our study, we found that the prevalence of PCOS in subjects without PCOS was 47.8%. This finding is similar to the results of other studies that have reported PCOS prevalence rates ranging from 9% to 33.9% in the general population [2,4,3].

Bottom line

We found no significant difference in the follicle to stroma ratio in ovarian biopsies from subjects without PCOS and those with PCOS. In subjects with PCOS, we found no significant difference in the follicle to stroma ratio among the follicle diameter group. However, a larger follicle size was associated with a higher follicle to stroma ratio.

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3. Reaven T. Pathogenesis of polycystic ovary syndrome. J Clin Endocrinol Metab. 2003;88(2):554556.
4. Matsuura Y, et al. Association between polycystic ovary syndrome and the metabolic syndrome in Japanese women: a hospital-based study. J Clin Endocrinol Metab. 2006;91(5):935.
5. Larian S, et al. Polycystic ovary syndrome: a review of the disease. Am J Obstet Gynecol. 2009;199(1):54.
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7. Girolami E, et al. Polycystic ovary syndrome: a review of its epidemiology and pathophysiology. Endocr Rev. 2013;32(4):382312.
8. Kim J, et al. Insulin resistance in polycystic ovary syndrome: a meta-analysis. Hum Biol. 2012;27(7):82231.

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