Skip to main content

Isolation of Human Gastric Epithelial Cells from Gastric Surgical Tissue and Gastric Biopsies for Primary Culture

  • Protocol
  • First Online:
Epithelial Cell Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1817))

Abstract

The epithelium of human gastric mucosa is involved in secretory and digestive functions, and distinct epithelial populations located in specific compartments of the gastric pit-gland units mediate these functions. We successfully developed a method for the isolation and growth of normal human gastric epithelial cells using biopsies or surgically resected tissues as the source of the cells. Gastric epithelial cell aggregates were released from the underlying tissues by gentle enzymatic digestion of collagenase type IV. Primary cultures were generated by seeding viable multicellular aggregates on plastic without a biological matrix. Characterized by immunostaining of cell-specific antigens, the cells were confirmed to be heterogenous gastric epithelial primary cultures containing mucus cells, neck cells, parietal cells, chief cells, and gastrin-secreting endocrine cells. This simple and convenient method will prove useful in the isolation of normal human gastric mucous epithelial cells for in vitro studies of gastric epithelial biology.

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

Access this chapter

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
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
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. Modlin IM, Kidd M, Lye KD et al (2003) Gastric stem cells: an update. Keio J Med 52(2):134–137

    Article  CAS  PubMed  Google Scholar 

  2. Karam SM, Straiton T, Hassan WM et al (2003) Defining epithelial cell progenitors in the human oxyntic mucosa. Stem Cells 21(3):322–336. https://doi.org/10.1634/stemcells.21-3-322

    Article  PubMed  Google Scholar 

  3. Sanders MJ, Amirian DA, Ayalon A et al (1983) Regulation of pepsinogen release from canine chief cells in primary monolayer culture. Am J Phys 245(5 Pt 1):G641–G646

    CAS  Google Scholar 

  4. Rattner DW, Ito S, Rutten MJ et al (1985) A rapid method for culturing guinea pig gastric mucous cell monolayers. In Vitro Cell Dev Biol 21(8):453–462

    Article  CAS  PubMed  Google Scholar 

  5. Chew CS (1994) Parietal cell culture: new models and directions. Annu Rev Physiol 56:445–461. https://doi.org/10.1146/annurev.ph.56.030194.002305

    Article  PubMed  CAS  Google Scholar 

  6. Terano A, Ivey KJ, Stachura J et al (1982) Cell culture of rat gastric fundic mucosa. Gastroenterology 83(6):1280–1291

    CAS  PubMed  Google Scholar 

  7. Terano A, Mach T, Stachura J et al (1983) A monolayer culture of human gastric epithelial cells. Dig Dis Sci 28(7):595–603

    Article  PubMed  CAS  Google Scholar 

  8. Rutten MJ, Campbell DR, Luttropp CA et al (1996) A method for the isolation of human gastric mucous epithelial cells for primary cell culture: a comparison of biopsy vs surgical tissue. Methods Cell Sci 18(4):269–281. https://doi.org/10.1007/bf00127904

    Article  Google Scholar 

  9. Moyer MP (1983) Culture of human gastrointestinal epithelial cells. Proc Soc Exp Biol Med 174(1):12–15

    Article  PubMed  CAS  Google Scholar 

  10. Quaroni A, May RJ (1980) Establishment and characterization of intestinal epithelial cell cultures. Methods Cell Biol 21B:403–427

    Article  PubMed  CAS  Google Scholar 

  11. Kedinger M, Haffen K, Simon-Assmann P (1987) Intestinal tissue and cell cultures. Differentiation 36(1):71–85

    Article  PubMed  CAS  Google Scholar 

  12. Evans GS, Flint N, Potten CS (1994) Primary cultures for studies of cell regulation and physiology in intestinal epithelium. Annu Rev Physiol 56:399–417. https://doi.org/10.1146/annurev.ph.56.030194.002151

    Article  PubMed  CAS  Google Scholar 

  13. Ito H, Clancy TE, Osteen RT et al (2004) Adenocarcinoma of the gastric cardia: what is the optimal surgical approach? J Am Coll Surg 199(6):880–886. https://doi.org/10.1016/j.jamcollsurg.2004.08.015

    Article  PubMed  Google Scholar 

Download references

Acknowledgement

This work was supported by the National Key Research and Development Program of China (2017YFA0103100, 2017YFA0103103, 2017YFA0103104), Guangdong Frontier and Key Technology Innovation Project (2014B020228001, 2014B020228003), and Guangzhou Health Care and Cooperative Innovation Major Project (201508020262, 201604020001, 201704020218).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Xuetao Pei .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Qin, J., Pei, X. (2018). Isolation of Human Gastric Epithelial Cells from Gastric Surgical Tissue and Gastric Biopsies for Primary Culture. In: Baratta, M. (eds) Epithelial Cell Culture. Methods in Molecular Biology, vol 1817. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8600-2_12

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8600-2_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8599-9

  • Online ISBN: 978-1-4939-8600-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics