siRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts.

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AuteurLOKOSSOU, ADJIMON GBENAKPON GATIEN
AuteurTOUFAILY, CHIRINE
AuteurVARGAS, AMANDINE
AuteurBARBEAU, BENOIT
Date d'ajout2026-06-02T16:06:57Z
Date de disponibilite2026-06-02T16:06:57Z
Date de publication2016
ResumeHuman primary villous cytotrophoblasts are a very useful source of primary cells to study placental functions and regulatory mechanisms, and to comprehend diseases related to pregnancy. In this protocol, human primary villous cytotrophoblasts freshly isolated from placentas through a standard DNase/trypsin protocol are microporated with small interfering RNA (siRNA). This approach provided greater efficiency for siRNA transfection when compared to a lipofection-based method. Transfected cells can subsequently be analyzed by standard Western blot within a time frame of 3-4 days post-transfection. In addition, using cultured primary villous cytotrophoblasts, Electrophoretic Mobility Shift Assay (EMSA) analysis was optimized and performed on extracts from days 1 to 4. The use of these cultured primary cells and the protocol described allow for an evaluation of the implication of specific genes and transcription factors in the process of villous cytotrophoblast differentiation into a syncytiotrophoblast-like cell layer. However, the limited time span allowable in culture precludes the use of methods requiring more time, such as generation of a stable cell population. Therefore testing of this cell population requires highly optimized gene transfer protocols.
DOI10.3791/53995
Autre identifiantBECDB-4117
URIhttps://dspace.uac.bj/handle/123456789/3946
Languefr
Fait partie deJournal of visualized experiments : JOVE
SujetCytotrophoblast
SujetsiRNA
Sujettransfection
SujetEMSA
TitresiRNA Transfection and EMSA Analyses on Freshly Isolated Human Villous Cytotrophoblasts.
TypeArticle

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