Syncytiotrophoblast cells are essential in placental development and function. Their successful generation is critical for various studies in reproductive biology and pregnancy-related research. This protocol outlines the materials required and a step-by-step procedure for differentiating pluripotent stem cells into syncytiotrophoblast cells.
Thaw and culture pluripotent stem cells on Matrigel-coated plates until they reach 70-80% confluency. When cells reach 70-80% confluence, split them using a gentle cell dissociation reagent to achieve a single-cell suspension.
Plate single-cell suspension onto Matrigel-coated dishes at a certain density. Cultivate pluripotent stem cells in DMEM/F12 medium supplemented with FBS, BMP4, Activin A, and FGF4. Change the medium every 24 hours for the first three days.
After three days of differentiation, observe the formation of trophoblast spheroids. Carefully collect spheroids by gentle pipetting.
Plate trophoblast spheroids onto fresh Matrigel-coated dishes. Maintain them in DMEM/F12 medium supplemented with FBS, BMP4, and FGF4. Change the medium every 48 hours.
After approximately one week of culture, syncytiotrophoblast-like cells should become evident. Confirm syncytiotrophoblast identity through immunofluorescence staining for markers. Harvest syncytiotrophoblast cells for further analysis or applications.
Creative Biolabs is dedicated to advancing stem cell research, and we are available to assist with any further inquiries or customization of this protocol to meet specific research needs.
Please feel free to contact our team for further assistance and support in your research endeavors.
Reference
For Research Use Only. Not For Clinical Use.
