Method for efficiently separating primary human endometrial stromal and epithelial cells

Objective To establish an efficient method for isolating primary human endometrium stromal and epithelial cells in vitro. Methods Health endometrial tissue of hysterectomy patient was obtained, and was cut into a small piece. Firstly, the human endometrial stromal cells (HUSC) and human endometrial epithelial cells (HUEC) were isolated by the twice enzymatic digestion, twice separation and different adherent time of the two types of cells. And then HUSC and HUEC were identified and the percentage of vimentin (Vim)-positive and cytokeratin (CK)-positive ones were determined by flow cytometry. After that, primary HUSC were stimulated by cyclic adenosine monophosphate (cAMP)+medroxyprogesterone acetate (MPA) for 4 d and 7 d. Results The HUSC purity was (97.0±2.5)% and the HUEC purity was (90.0±4.1)% determined by flow cytometry, indicating that we had highly purified cultures of HUSC and HUEC in vitro. After cAMP+MPA induction for 4 d and 7 d, the HUSC showed more obvious changes in cell morphology and prolactin (PRL) level. Conclusion We successfully established an efficient method of isolation and characterization of primary human endometrium stromal and epithelial cells in vitro. Key words: Human endometrium stromal cells; Human endometrium epithelial cells; Isolation method; Molecular markers; Decidualization