Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell specific gene expression and cytokine production

Capsule:
A coculture system of polarized human endometrial epithelium and subject-paired stroma reveals effects of paracrine signaling on cell-specific global gene expression and cytokine production.

Authors:
Joseph C. Chen, Ph.D., M.S., David W. Erikson, Ph.D., Terhi T. Piltonen, M.D., Ph.D., Michelle R. Meyer, B.S., Fatima Barragan, B.S., Ramsey H. McIntire, Ph.D., John S. Tamaresis, Ph.D., Kim Chi Voa, B.S., Linda C. Giudice, M.D., Ph.D., M.Sc., Juan C. Irwina, M.D., Ph.D.

Volume 100, Issue 4, Pages 1132-1143, October 2013

Abstract:

Objective:
To determine the effects of coculturing endometrial epithelial cells (eEC) with paired endometrial stromal fibroblasts (eSF) on cell-specific gene expression and cytokine secretion patterns.

Design:
In vitro study.

Setting:
University research laboratory.

Patient(s):
Endometrial biopsies were obtained from premenopausal women.

Intervention(s):
Polarized eEC and subject-paired eSF were cultured for 12.5 hours alone (monoculture) or combined in a two-chamber coculture system without cell-cell contact. Cells and conditioned media were analyzed for global gene expression and cytokine secretion, respectively. Purified, endometrial tissue-derived eEC and eSF isolated by fluorescent activated cell sorting (FACS) were used as noncultured controls.

Main Outcome Measure(s):
Cell-specific global gene expression profiling and analysis of secreted cytokines in eEC/eSF cocultures and respective monocultures.

Result(s):
Transepithelial resistance, diffusible tracer exclusion, expression of tight junction proteins, and apical/basolateral vectorial secretion confirmed eEC structural and functional polarization. Distinct transcriptomes of eEC and eSF were consistent with their respective lineages and their endometrial origin. Coculture of eEC with eSF resulted in altered cell-specific gene expression and cytokine secretion.

Conclusion(s):
This coculture model provides evidence that interactions between endometrial functionally polarized epithelium and stromal fibroblasts affect cell-specific gene expression and cytokine secretion underscoring their relevance when modeling endometrium in vitro.

  • Stefania Salsano

    Dear Dr. Chen, can you give me an email address please? I’ve some technical questions! Thank you

    • Joe C

      Dear Stefania,
      I’m sorry I did not check this website for queries. You can email me at joseph.chen@ucsf.edu.
      Sorry for the late response!

      • Joe–

        please post the technical details here for everyone to learn from!

        Steven Palter, MD
        New Media Editor
        Fertility Sterility

        • Joe C

          OK will do. If Stefania contacts me I will post the question and the specific response on the technical issue. Thank you. And I shall check back here more regularly.

    • stefania—could you please share your technical question here instead of email so everyone can learn together! thanks

      Steven Palter, MD
      New Media Editor
      Fertility Sterility

  • Jacquelyn Hoffman

    This is great! Question for you, Dr. Chen: are these epithelial cells able to be frozen and banked?

    • Joe Chen

      Not yet but currently we are trying to establish those cryo-preservation protocols so that these cells can be sent to other researchers for experimentation. If you have any suggestions Jacquelyn, from your banking experience, would love to hear it!

  • Joe Chen

    Please contact me for any technical details, thank you!

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