Isolation of human endometrial stromal cells for in vitro decidualization

Stephanie A. Michalski, Sangappa B. Chadchan, Emily S. Jungheim, Ramakrishna Kommagani*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


The differentiation of human endometrial stromal cells (HESC) from fibroblast-like appearance into secretory decidua is a transformation required for embryo implantation into the uterine lining of the maternal womb. Improper decidualization has been established as a root cause for implantation failure and subsequent early embryo miscarriage. Therefore, understanding the molecular mechanisms underlying decidualization is advantageous to improving the rate of successful births. In vivo based studies of artificial decidualization are often limiting due to ethical dilemmas associated with human research, as well as translational complications within animal models. As a result, in vitro assays through primary cell culture are often utilized to explore the modulation of decidualization via hormones. This study provides a detailed protocol for the isolation of HESC and subsequent artificial decidualization via the supplementation of hormones to the culturing medium. Further, this study provides a well-designed method to knockdown any gene of interest by utilizing lipid-based siRNA transfections. This protocol permits the optimization of culture purity as well as product yield, thereby maximizing the ability to utilize this model as a reliable method to understand the molecular mechanisms underlying decidualization, and the subsequent quantification of secreted agents by decidualized endometrial stromal cells.

Original languageEnglish (US)
Article numbere57684
JournalJournal of Visualized Experiments
Issue number139
StatePublished - Sep 1 2018
Externally publishedYes


  • And in vitro decidualization
  • Camp
  • Developmental biology
  • Developmental biology
  • Estrogen
  • Human endometrial stromal cells
  • Issue 139
  • Primary cultures
  • Progesterone
  • Sirna transfection

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)


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