complete expansion medium  (ATCC)

Journal: Cell Death & Disease

Article Title: CRISPR/Cas9-mediated gene editing in trophoblast cells via mechanoporation for preeclampsia insight

doi: 10.1038/s41419-025-08200-z

Figure Lengend Snippet: sEVs were isolated from MSCs and labeled with PKH67 prior to incubation with ACH-3P cells. Cells were incubated for 2, 4 and 6 h prior to washing and fixing. Untreated cells were used as a negative control (NC). Hoechst 33342 was used to label nuclei. A Representative maximum intensity projection images. Images were acquired at 20× magnification using a Leica Stellaris confocal microscope. Scale bar = 50 µm. B Fluorescent intensity of PKH67-labeled sEVs internalized within trophoblast cells across incubation timepoints. Data expressed as mean ± SEM, n = 3. C High-resolution image acquired at 100× magnification using a Nikon A1R confocal microscope. The scale bar indicates 20 µm. D WT, K/O15 and K/O23 cells were treated with PKH67-labeled sEVs for 2, 4, and 6 h. Images were acquired with a Zeiss Lattice SIM microscope. Scale bar = 50 µm. E Mean fluorescent intensity of sEVs in cells was analyzed using ImageJ and normalized to the WT control. NC negative control, WT wildtype, K/O knockout, sEVs small extracellular vesicles.

Article Snippet: Primary bone marrow-derived mesenchymal stem cells (BM-MSCs; ATCC, United States) were initially expanded in ATCC mesenchymal stem cell basal medium and growth kit (ATCC, United States) prior to maintenance in Dulbecco’s Modified Eagle’s Medium (DMEM)/F12 + GlutaMAX (Thermo Fisher Scientific, United States) with 10% FBS and 1% P/S.

Techniques: Isolation, Labeling, Incubation, Negative Control, Microscopy, Control, Knock-Out