Skip to main content
SpringerLink
Log in

A Simplified Method for Three-Dimensional (3D) Porcine Preantral Follicles Culture Utilizing Hydrophobic Microbioreactors

  • Protocol
  • First Online:
Book cover Next Generation Culture Platforms for Reliable In Vitro Models

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2273))

  • 1163 Accesses

Abstract

The technological revolution in reproductive biology that started with artificial insemination procedures and embryo transfer led to the development of assisted reproduction techniques such as in vitro fertilization or even cloning of domestic animals by nuclear transfer from somatic cells. Currently, procedures of isolated immature ovarian follicles in vitro culture are becoming the prominent technology aimed to preserve or restore fertility especially of young oncological patients or those at risk of premature ovarian failure.

Here, we describe a protocol that can be applied for in vitro growth of porcine, preantral ovarian follicles in three-dimensional (3D) culture conditions. After enzymatic isolation from the ovarian cortex, preantral follicles are suspended in a drop of medium and enclosed with fluorinated ethylene propylene (FEP) powder particles (microbioreactors). Such microbioreactors maintain the 3D structure of the follicles during the whole process of in vitro growth what is crucial to ensure proper folliculogenesis progression and their ability to survive.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Fortune JE, Ribera GM, Yang MY (2004) Follicular development: the role of follicular microenvironment in selection of dominant follicle. Anim Reprod Sci 82-83:109–126. https://doi.org/10.1016/j.anireprosci.2004.04.031

    Article  CAS  PubMed  Google Scholar 

  2. Telfer EE, Binnie JP, McCaffery FH et al (2000) In vitro development of oocytes from porcine and bovine primary follicles. Mol Cell Endocrinol 163:117–123. https://doi.org/10.1016/s0303-7207(00)00216-1

    Article  CAS  PubMed  Google Scholar 

  3. Wu J, Emery BR, Carrell DT (2001) In vitro growth, maturation, fertilization, and embryonic development of oocytes from porcine preantral follicles. Biol Reprod 64:375–381. https://doi.org/10.1095/biolreprod64.1.375

    Article  CAS  PubMed  Google Scholar 

  4. Abir R, Nitke S, Ben-Haroush A et al (2006) In vitro maturation of human primordial ovarian follicles: clinical significance, progress in mammals, and methods for growth evaluation. Histol Histopathol 21:887–898. https://doi.org/10.14670/HH-21.887

    Article  CAS  PubMed  Google Scholar 

  5. Kreeger PK, Deck JW, Woodruff TK et al (2006) The in vitro regulation of ovarian follicle development using alginate-extracellular matrix gels. Biomaterials 27:714–723. https://doi.org/10.1016/j.biomaterials.2005.06.016

    Article  CAS  PubMed  Google Scholar 

  6. Eppig JJ, Schroeder AC (1989) Capacity of mouse oocytes from preantral follicles to undergo embryogenesis and development to live young after growth, maturation, and fertilization in vitro. Biol Reprod 41:268–276. https://doi.org/10.1095/biolreprod41.2.268

    Article  CAS  PubMed  Google Scholar 

  7. Eppig JJ, O’Brien MJ (1996) Development in vitro of mouse oocytes from primordial follicles. Biol Reprod 54:197–207. https://doi.org/10.1095/biolreprod54.1.197

    Article  CAS  PubMed  Google Scholar 

  8. Oktem O, Oktay K (2007) Sphingosine-1-phosphate enhances human primordial follicle survival and blocks ovarian apoptosis in vitro. Paper presented at the ASRM 63rd Annual Meeting, Washington, DC, USA 15–17 Oct 2007

    Google Scholar 

  9. Belli M, Vigone G, Merico V et al (2012) (2012) towards a 3D culture of mouse ovarian follicles. Int J Dev Biol 56:931–937. https://doi.org/10.1387/ijdb.120175mz

    Article  CAS  PubMed  Google Scholar 

  10. Desai N, Alex A, Abdel Hafez F et al (2010) Three-dimensional in vitro follicle growth: overview of culture models, biomaterials, design parameters and future directions. Reprod Biol Endocrinol 8:119. https://doi.org/10.1186/1477-7827-8-119

    Article  PubMed  PubMed Central  Google Scholar 

  11. Filatov MA, Khramova YV, Semenova ML (2015) In vitro mouse ovarian follicle growth and maturation in alginate hydrogel: current state of the art. Acta Nat 7:48–56

    Article  CAS  Google Scholar 

  12. Abir R, Fisch B, Nitke S et al (2001) Morphological study of fully and partially isolated early human follicles. Fertil Steril 75:141–146. https://doi.org/10.1016/s0015-0282(00)01668-x

    Article  CAS  PubMed  Google Scholar 

  13. Cukierman E, Pankov R, Yamada KM (2002) Cell interactions with three-dimensional matrices. Curr Opin Cell Biol 14:633–639. https://doi.org/10.1016/s0955-0674(02)00364-2

    Article  CAS  PubMed  Google Scholar 

  14. Combelles CMH, Fissore RA, Albertini DF et al (2005) In vitro maturation of human oocytes and cumulus cells using a co-culture three-dimensional collagen gel system. Hum Reprod 20:1349–1358. https://doi.org/10.1093/humrep/deh750

    Article  CAS  PubMed  Google Scholar 

  15. Dorati R, Genta I, Ferrari M et al (2016) Formulation and stability evaluation of 3D alginate beads potentially useful for cumulus-oocyte complexes culture. J Microencapsul 33:137–145. https://doi.org/10.3109/02652048.2015.1134691

    Article  CAS  PubMed  Google Scholar 

  16. Sadr SZ, Fatehi R, Maroufizadeh S et al (2018) Utilizing fibrin-alginate and matrigel-alginate for mouse follicle development in three-dimensional culture systems. Biopreserv Biobank 16(2):120–127. https://doi.org/10.1089/bio.2017.0087

    Article  CAS  PubMed  Google Scholar 

  17. Pangas SA, Saudye H, Shea LD et al (2003) Novel approach for the three-dimensional culture of granulosa cell-oocyte complexes. Tissue Eng 9:1013–1021. https://doi.org/10.1089/107632703322495655

    Article  CAS  PubMed  Google Scholar 

  18. Xu M, West E, Shea LD et al (2006) Identification of a stage-specific permissive in vitro culture environment for follicle growth and oocyte development. Biol Reprod 75:916–923. https://doi.org/10.1095/biolreprod.106.054833

    Article  CAS  PubMed  Google Scholar 

  19. West ER, Xu M, Woodruff TK et al (2007) Physical properties of alginate hydrogels and their effects on in vitro follicle development. Biomaterials 28:4439–4448. https://doi.org/10.1016/j.biomaterials.2007.07.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Xu M, Kreeger PK, Shea LD et al (2006) Tissue-engineered follicles produce live, fertile offspring. Tissue Eng 12:2739–2746. https://doi.org/10.1089/ten.2006.12.2739

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Jin SY, Lei L, Shikanov A et al (2010) A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse. Fertil Steril 93:2633–2639. https://doi.org/10.1016/j.fertnstert.2009.10.027

    Article  PubMed  Google Scholar 

  22. Parrish EM, Siletz A, Xu M et al (2011) Gene expression in mouse ovarian follicle development in vivo versus an ex vivo alginate culture system. Reproduction 142:309–318. https://doi.org/10.1530/REP-10-0481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Arbatan T, Al-Abboodi A, Sarvi F et al (2012) Tumor inside a pearl drop. Adv Healthc Mater 1:467–469. https://doi.org/10.1002/adhm.201200050

    Article  CAS  PubMed  Google Scholar 

  24. Tian J, Fu N, Chen XD et al (2013) Respirable liquid marble for the cultivation of microorganisms. Colloids Surf B Biointerfaces 106:187–190. https://doi.org/10.1016/j.colsurfb.2013.01.016

    Article  CAS  PubMed  Google Scholar 

  25. Sarvi F, Jain K, Arbatan T et al (2015) Cardiogenesis of embryonic stem cells with liquid marble micro-bioreactor. Adv Healthc Mater 4(1):77–86. https://doi.org/10.1002/adhm.201400138

    Article  CAS  PubMed  Google Scholar 

  26. Ledda S, Idda A, Kelly J et al (2016) A novel technique for in vitro maturation of sheep oocytes in a liquid marble microbioreactor. J Assist Reprod Genet 33:513–518. https://doi.org/10.1007/s10815-016-0666-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Gorczyca G, Wartalski K, Tabarowski Z et al (2020) Proteolytically degraded alginate hydrogels and hydrophobic microbioreactors for porcine oocyte encapsulation. J Vis Exp. https://doi.org/10.3791/61325

Download references

Acknowledgments

The authors are very grateful to Dr. Zbigniew Tabarowski (Department of Experimental Hematology, Institute of Zoology and Biomedical Research, Jagiellonian University) for critical revision of the manuscript. This work was supported by grant 2018/29/N/NZ9/00983 from National Science Centre Poland. GG and MD are members of the COST Actions CA16119.

Author information

Authors and Affiliations

  1. Department of Endocrinology, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University in Krakow, Kraków, Poland

    Malgorzata Duda, Lucia Gizler & Gabriela Gorczyca

Authors
  1. Malgorzata Duda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lucia Gizler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gabriela Gorczyca
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Malgorzata Duda .

Editor information

Editors and Affiliations

  1. Laboratory of Biomedical Embryology, Università degli Studi di Milano, Milan, Italy

    Tiziana A.L. Brevini

  2. Department of Pathophysiology, University of Tartu, Tartu, Estonia

    Alireza Fazeli

  3. Ottawa, ON, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2021 The Author(s), under exclusive license to Springer Science+Business Media, LLC , part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Duda, M., Gizler, L., Gorczyca, G. (2021). A Simplified Method for Three-Dimensional (3D) Porcine Preantral Follicles Culture Utilizing Hydrophobic Microbioreactors. In: Brevini, T.A., Fazeli, A., Turksen, K. (eds) Next Generation Culture Platforms for Reliable In Vitro Models . Methods in Molecular Biology, vol 2273. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1246-0_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1246-0_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1245-3

  • Online ISBN: 978-1-0716-1246-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation