Category Archives: Human Placenta

Expression of the Erythropoietin Receptor: MATERIALS AND METHODS(3)

The procedure for trophoblast isolation was based on the method of Kliman et al. with modifications. Briefly, the basal plate was removed, and approximately 50 g of villous tissue was harvested and teased into small fragments. Villous trophoblast cells were released by four sequential trypsin digestions (trypsin grade III; Sigma Chemical Co., St. Louis, MO). DNase I (grade II; Boehringer Mannheim, Indianapolis, IN) was used to prevent cellular aggregation. The first digestion was routinely discarded, as it contained few trophoblast cells and mainly red blood cells and leukocytes. The digests were layered over FBS and centrifuged to remove cellular debris and inactivate trypsin. antibiotic levaquin

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