TY - JOUR
T1 - Isolation of human endometrial stromal cells for in vitro decidualization
AU - Michalski, Stephanie A.
AU - Chadchan, Sangappa B.
AU - Jungheim, Emily S.
AU - Kommagani, Ramakrishna
N1 - Publisher Copyright:
© 2018, Journal of Visualized Experiments. All rights reserved.
PY - 2018/9/1
Y1 - 2018/9/1
N2 - 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.
AB - 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.
KW - And in vitro decidualization
KW - Camp
KW - Developmental biology
KW - Developmental biology
KW - Estrogen
KW - Human endometrial stromal cells
KW - Issue 139
KW - Primary cultures
KW - Progesterone
KW - Sirna transfection
UR - http://www.scopus.com/inward/record.url?scp=85054376538&partnerID=8YFLogxK
U2 - 10.3791/57684
DO - 10.3791/57684
M3 - Article
C2 - 30222162
AN - SCOPUS:85054376538
SN - 1940-087X
VL - 2018
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 139
M1 - e57684
ER -