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Characterization of stem cells from human ovarian follicular fluid; a potential source of autologous stem cell for cell-based therapy

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Abstract

Human ovarian follicular fluid (HOFF) contains proteins, extracellular matrixes necessary for growth and maturation of oocytes as well as granulosa cells. Epithelial cells and stem cells can be isolated from HOFF. However, information regarding stem cells derived from HOFF is still lacking. The objectives of the present study were to isolate, characterize, and differentiate cells derived from HOFF. HOFF was collected during the routine aspiration of oocytes in an assisted fertilization program and subjected to cell isolation, characterization, and in vitro culture. After 24 h of culture, different cell morphologies including epithelial-like-, neural-like- and fibroblast-like cells were observed. Immunocytochemistry reveals the expression of pluripotent stem cell markers (OCT4, NANOG, SSEA4), epithelial marker (CK18), FSH- and LH-receptor. For in vitro culture, the isolated cells were continuously cultured in a growth medium; alpha MEM containing 10% FBS and epidermal growth factor (EGF). After 2 weeks of in vitro culture, cells with fibroblast-like morphology dominantly grow in the culture vessels and resemble mesenchymal stem cells (MSCs). HOFF-derived cells exhibited MSC expression of CD44, CD73, CD90, CD105, CD146, and STRO-1, and were capable of differentiation into osteoblasts, chondrocytes, and adipocytes. After induction of neural differentiation, HOFF-derived cells formed spheroidal structures and expressed neural stem cell markers including Nestin, β-tubulin III, and O4. Besides, the oocyte-like structure was observed after prolonged culture of HOFF. In conclusion, cells derived from follicular fluid exhibited stem cell characteristics, which could be useful for regenerative medicine applications and cell-based therapies.

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Acknowledgements

The present study was financially supported by the grant from Chulalongkorn University and Srinakharinwirot University (MEDRES 200). The authors are grateful to Dr. Carol Lynn Curchoe for the English editing of the manuscript.

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Authors and Affiliations

  1. Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand

    Ruttachuk Rungsiwiwut

  2. Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

    Pranee Numchaisrika, Paweena Thuwanut & Kamthorn Pruksananonda

Authors
  1. Ruttachuk Rungsiwiwut
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  2. Pranee Numchaisrika
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  3. Paweena Thuwanut
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  4. Kamthorn Pruksananonda
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Correspondence to Kamthorn Pruksananonda.

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The authors declare no conflicts of interest.

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The protocol of the present study was approved by the Ethical Committee of the Faculty of Medicine, Chulalongkorn University.

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The informed consent was obtained from all individual participants included in the present study.

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Rungsiwiwut, R., Numchaisrika, P., Thuwanut, P. et al. Characterization of stem cells from human ovarian follicular fluid; a potential source of autologous stem cell for cell-based therapy. Human Cell 34, 300–309 (2021). https://doi.org/10.1007/s13577-020-00439-2

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  • DOI: https://doi.org/10.1007/s13577-020-00439-2

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