Stem Cell Reviews and Reports

, Volume 15, Issue 4, pp 543–557 | Cite as

Female Age Affects the Mesenchymal Stem Cell Characteristics of Aspirated Follicular Cells in the In Vitro Fertilization Programme

  • Irma Virant-KlunEmail author
  • S. Omejec
  • M. Stimpfel
  • P. Skerl
  • S. Novakovic
  • N. Jancar
  • E. Vrtacnik-Bokal


Aspirated follicular cells (AFCs) from the in vitro fertilization program can express various stem cell markers and are even able to differentiate into different types of cells in vitro. The female reproductive potential decreases with increasing age due to lowered ovarian reserve and oocyte quality, but data on the effect of female age on stem cell characteristics of AFCs are scarce. Therefore, the aim of this study was to elucidate whether female age affects the mesenchymal stem cell (MSC) characteristics of AFCs. Follicular aspirates were collected from 12 patients included in the in vitro fertilization programme with a normal ovarian reserve. Patients were divided into four age groups: Group A ≤ 30 years, Group B 31–35 years, Group C 36–39 years and Group D ≥ 40 years. After removal of the oocytes, AFCs were collected from follicular aspirates using hypo-osmotic technique and cultured in vitro, and their stemness was compared according to female age. The cultured AFCs were analysed for gene expression using the Human Mesenchymal Stem Cell RT2 Profiler™ PCR Array, for their potential for differentiation into adipogenic and osteogenic lineage, and for their expression of MSC-related markers using immunocytochemistry. We found that female age can significantly influence their stemness: expression of pluripotency and MSC-related genes, and their differentiation potential. Despite the relatively high expression of MSC-related genes, the AFCs of the oldest patients had the lowest potential to differentiate into osteogenic and adipogenic lineages in vitro, which may be related to their age and the changed ovarian function.


Aspirated follicular cells Granulosa cells Mesenchymal stem cells Female age Stem cells Differentiation 



The authors thank all of our colleagues at the Reproductive Unit, Department of Obstetrics and Gynaecology, University Medical Centre Ljubljana for their collaboration and support and all of the patients who kindly donated their follicular fluid for this research.

Authors’ Contributions

IVK designed the experiments, organized the genetic analysis, interpreted the results, wrote the manuscript, and financially supported the work by her research project; SO performed the experimental work, collected the data and wrote the manuscript; MS performed the experimental work and wrote the manuscript; EVB and NJ performed the ultrasound-guided aspiration of follicular fluid and discussed the clinical part of the work; PS and SN performed the genetic analysis of AFCs, discussed the interpretation of genetic data, and read and corrected the manuscript.


The research was funded by the research grant J3–4195 offered to Prof. Irma Virant-Klun by the Slovenian Research Agency (ARRS).

Compliance with Ethical Standards

Conflict of Interest

The authors have no financial or other conflicts of interests about this work to declare.

Supplementary material

12015_2019_9889_MOESM1_ESM.doc (158 kb)
ESM 1 (DOC 158 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Irma Virant-Klun
    • 1
    Email author
  • S. Omejec
    • 2
  • M. Stimpfel
    • 1
  • P. Skerl
    • 3
  • S. Novakovic
    • 3
  • N. Jancar
    • 1
  • E. Vrtacnik-Bokal
    • 1
  1. 1.Department of Obstetrics and GynaecologyUniversity Medical Centre LjubljanaLjubljanaSlovenia
  2. 2.Blood Transfusion Centre of SloveniaLjubljanaSlovenia
  3. 3.Department of Molecular DiagnosticsInstitute of Oncology LjubljanaLjubljanaSlovenia

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