Skip to main content
SpringerLink
Log in

Isolation of Normal Fibroblasts and Their Cancer-Associated Counterparts (CAFs) for Biomedical Research

  • Protocol
  • First Online:
Skin Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1879))

Abstract

Cancer-associated fibroblasts (CAFs) represent a crucial component of cancer microenvironment. CAFs significantly influence biological properties of various types of cancer in terms of local aggressiveness, recurrence, and metastatic behaviour. This chapter summarizes a simple protocol for isolation of normal fibroblasts and their cancer-associated counterparts from normal human skin and mucosa, respectively, as well as from samples of human tumours such as basal/squamous carcinoma, melanoma, and breast cancer, and employment of this procedure for other types of cancer is possible. Isolated fibroblasts can be expanded in vitro and employed for further analysis of, e.g., DNA, RNA, protein, etc.

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 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 169.99
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. Marks PW, Witten CM, Califf RM (2017) Clarifying stem-cell therapy’s benefits and risks. N Engl J Med 376:1007–1009

    Article  Google Scholar 

  2. Caplan AI (2017) Mesenchymal Stem Cells: Time to Change the Name! Stem Cells Transl Med 6:1445–1451

    Article  Google Scholar 

  3. Bardelli S, Moccetti M (2017) Remodeling the human adult stem cell niche for regenerative medicine applications. Stem Cells Int 2017:6406025

    PubMed  PubMed Central  Google Scholar 

  4. Madl CM, Heilshorn SC (2017) Engineering hydrogel microenvironments to recapitulate the stem cell niche. Annu Rev Biomed Eng. https://doi.org/10.1146/annurev-bioeng-062117-120954

    Article  CAS  Google Scholar 

  5. Szade KT, Gulati GS, Chan CKF et al (2017) Where hematopoietic stem cells live—the bone marrow niche. Antioxid Redox Signal. https://doi.org/10.1089/ars.2017.7419

    Article  CAS  Google Scholar 

  6. Machida K (2017) Pluripotency transcription factors and metabolic reprogramming of mitochondria in tumor-initiating stem-like cells. Antioxid Redox Signal. https://doi.org/10.1089/ars.2017.7241

    Article  CAS  Google Scholar 

  7. Motlík J, Klíma J, Dvoránková B, Smetana K Jr (2007) Porcine epidermal stem cells as a biomedical model for wound healing and normal/malignant epithelial cell propagation. Theriogenology 67:105–111

    Article  Google Scholar 

  8. Kareva I (2011) What can ecology teach us about cancer? Transl Oncol 4:266–270

    Article  Google Scholar 

  9. Kareva I (2013) Biological stoichiometry in tumor micro-environments. PLoS One 8:e51844

    Article  CAS  Google Scholar 

  10. Lacina L, Plzak J, Kodet O et al (2015) Cancer microenvironment: what can we learn from the stem cell niche. Int J Mol Sci 16:24094–24110

    Article  Google Scholar 

  11. Gál P, Varinská L, Fáber L et al (2017) How signaling molecules regulate tumor microenvironment: parallels to wound repair. Molecules 22:E1818

    Article  Google Scholar 

  12. Bai X (ed) (2013) Dermal fibroblasts: histological perspectives, characterization and role in disease. Nova Sciences, New York

    Google Scholar 

  13. Živicová V, Lacina L, Mateu R et al (2017) Analysis of dermal fibroblasts isolated from neonatal and child cleft lip and adult skin: developmental implications on reconstructive surgery. Int J Mol Med 40:1323–1334

    Article  Google Scholar 

  14. Valach J, Fík Z, Strnad H et al (2012) Smooth muscle actin-expressing stromal fibroblasts in head and neck squamous cell carcinoma: increased expression of galectin-1 and induction of poor prognosis factors. Int J Cancer 131:2499–2508

    Article  CAS  Google Scholar 

  15. Trylcova J, Busek P, Smetana K Jr et al (2015) Effect of cancer-associated fibroblasts on the migration of glioma cells in vitro. Tumour Biol 36: 5873–5879

    Article  CAS  Google Scholar 

  16. Jobe NP, Rösel D, Dvořánková B et al (2016) Simultaneous blocking of IL-6 and IL-8 is sufficient to fully inhibit CAF-induced human melanoma cell invasiveness. Histochem Cell Biol 146:205–217

    Article  CAS  Google Scholar 

  17. Lacina L, Kodet O, Dvořánková B, Szabo P, Smetana K Jr (2018) Ecology of melanoma cell. Histol Histopathol 33:247–254

    CAS  PubMed  Google Scholar 

  18. Dvořánková B, Smetana K Jr, Říhová B, Kučera J, Mate R, Szabo P (2015) Cancer-associated fibroblasts are not formed from cancer cells by epithelial-to-mesenchymal transition in nu/nu mice. Histochem Cell Biol 143: 463–469

    Article  Google Scholar 

  19. Smetana K Jr, Lacina L, Szabo P, Dvořánková B, Brož P, Šedo A (2016) Ageing as an important risk factor for cancer. Anticancer Res 36:5009–5017

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This chapter was supported by the Grant Agency of the Czech Republic (grant no. 16-05534S); Charles University (project of PROGRES Q28/F1); Ministry of Education, Youth, and Sports of CR within the National Sustainability Program II (project BIOCEV-FAR; registration no. LQ1604); and project BIOCEV (grant no. CZ.1.05/1.1.00/02.0109). The present study is a result of the following project implementation: the equipment for metabolomic and cell analyses (grant no. CZ.1.05/2.1.00/19.0400) supported by the Research and Development for Innovations Operational Program, co-financed by the European regional development fund and the state budget of the Czech Republic.

Author information

Authors and Affiliations

  1. First Faculty of Medicine, Institute of Anatomy and BIOCEV, Charles University, Prague, Czech Republic

    BarboraBarbora Dvořánková, Lukáš Lacina & Karel Smetana Jr.

Authors
  1. BarboraBarbora Dvořánková
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lukáš Lacina
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Karel Smetana Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lukáš Lacina .

Editor information

Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media New York

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Dvořánková, B., Lacina, L., Smetana, K. (2018). Isolation of Normal Fibroblasts and Their Cancer-Associated Counterparts (CAFs) for Biomedical Research. In: Turksen, K. (eds) Skin Stem Cells. Methods in Molecular Biology, vol 1879. Humana Press, New York, NY. https://doi.org/10.1007/7651_2018_137

Download citation

  • DOI: https://doi.org/10.1007/7651_2018_137

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8869-3

  • Online ISBN: 978-1-4939-8870-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation