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Encapsulation of Mesenchymal Stem Cells in 3D Ovarian Cell Constructs Promotes Stable and Long-Term Hormone Secretion with Improved Physiological Outcomes in a Syngeneic Rat Model

  • Bioengineering and Enabling Technologies
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Abstract

Loss of ovarian function (e.g., due to menopause) leads to profound physiological effects in women including changes in sexual function and osteoporosis. Hormone therapies are a known solution, but their use has significantly decreased due to concerns over cardiovascular disease and certain cancers. We recently reported a tissue-engineering strategy for cell hormone therapy (cHT) in which granulosa cells and theca cells are encapsulated to mimic native ovarian follicles. cHT improved physiological outcomes and safety compared to pharmacological hormone therapies in a rat ovariectomy model. However, cHT did not achieve estrogen levels as high as ovary-intact animals. In this report, we examined if hormone secretion from cHT constructs is impacted by incorporation of bone marrow-derived mesenchymal stem cells (BMSC) since these cells contain regulatory factors such as aromatase necessary for estrogen production. Incorporation of BMSCs led to enhanced estrogen secretion in vitro. Moreover, cHT constructs with BMSCs achieved estrogen secretion levels significantly greater than constructs without BMSCs in ovariectomized rats from 70 to 90 days after implantation, while also regulating pituitary hormones. cHT constructs with BMSC ameliorated estrogen deficiency-induced uterine atrophy without hyperplasia. The results indicate that inclusion of BMSC in cHT strategies can improve performance.

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Acknowledgments

This study was supported, in part, by Jack and Pamela Egan. The study was also supported, in part, by the National Institutes of Health (NIH) Grant # R01AR061391 (JMS). The content of the article is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

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Authors and Affiliations

  1. Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, 27157, USA

    Sivanandane Sittadjody, Kevin M. Enck, Alexandra Wells, James J. Yoo, Anthony Atala & Emmanuel C. Opara

  2. Virginia Tech-Wake Forest School of Biomedical Engineering & Sciences (SBES), Wake Forest School of Medicine, Winston-Salem, NC, 27157, USA

    Kevin M. Enck, James J. Yoo, Anthony Atala & Emmanuel C. Opara

  3. Department of Chemical, Paper and Biomedical Engineering, Miami University, Oxford, OH, 45056, USA

    Justin M. Saul

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  1. Sivanandane Sittadjody
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Correspondence to Justin M. Saul.

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10439_2019_2334_MOESM1_ESM.tif

Supplemental Figure 1. Schematic of process of steroidogenesis. Theca cells express CYP11A1, 3βHSD, and CYP17A1, which are required for the production of androgens. Granulosa cells express aromatase, which allows for the conversion of androgens to estrone and estradiol. Thus, in native physiology, conversion of cholesterol to estrogen requires each of these enzymes. BMSCs express CYP19A1, but do not express CYP17A1, CYP11A1, or 3βHSD, which explains ability to enhance estrogen production but not progesterone. (TIFF 1405 kb)

10439_2019_2334_MOESM2_ESM.tif

Supplemental Figure 2. Additional immunohistochemistry for BMSCs. Cells are positive for CD105 (A). BMSC were negative for CD31 (B), CD45 (C), CYP17A1 (D), CYP11A1 (E), and 3βHSD (F). Positive staining for CD105 and negative staining for CD31 and CD45 are further evidence of isolation of BMSCs. The negative staining for CYP17A1, CYP11A1 and 3βHSD help to explain why the BMSCs are unable to produce progesterone and androgens. (TIFF 3947 kb)

10439_2019_2334_MOESM3_ESM.tif

Supplemental Figure 3. Resected omentum pouch after the in vivo treatment period of 90 days. Similar vascular density visible among the omentum pouches containing constructs with BMSC (A) and without BMSC (C). Gross images display intact microcapsule-constructs through the translucent omentum pouch containing constructs with BMSC (B) and without BMSC (D). The histology of resected omentum pouch shows no visible fibrous capsule around the constructs with BMSC (E) or without BMSC (F) (scale bar on E and F = 100 µm). (TIFF 13577 kb)

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Sittadjody, S., Enck, K.M., Wells, A. et al. Encapsulation of Mesenchymal Stem Cells in 3D Ovarian Cell Constructs Promotes Stable and Long-Term Hormone Secretion with Improved Physiological Outcomes in a Syngeneic Rat Model. Ann Biomed Eng 48, 1058–1070 (2020). https://doi.org/10.1007/s10439-019-02334-w

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