stemdiff apel 2 medium  (STEMCELL Technologies Inc)

Journal: Journal of Translational Medicine

Article Title: Interpenetrating network hydrogel-loaded embryonic stem cell-derived endocardial cells improves cardiac function after myocardial infarction

doi: 10.1186/s12967-025-06603-2

Figure Lengend Snippet: Biocompatibility of the hydrogel delivery system. A : ECCs were loaded in the hydrogel delivery system and prepared into frozen sections, and then immunofluorescence staining was performed on the frozen sections and observed under a confocal microscope (×10, CD31 + = the membrane of hESC-ECCs and red fluorescence + = hydrogel); B : SEM images (×500) of the hESC-ECCs loaded in the hydrogel delivery system (red cycles represent hESC-ECCs); C : Live/dead staining images of hESC-ECCs after culture in the hydrogel delivery system for 3, 14, and 28 d in LI-APEL medium in vitro (×5, green represents living cells, red represents dead cells); D : Calcein-AM staining was performed on the cell-loaded hydrogel after 3 d of culture to observe the distribution of hESC-ECCs (×5); E : Cck-8 detection was performed on the cell-loaded precrosslinking solution to test the cell numbers in different patches ( n = 5, ns P > 0.05). The data are shown as the means ± SDs, and three-group comparisons were performed via one-way ANOVA

Article Snippet: For CM and EC differentiation, hESCs (Cellapy) were treated with LI-APEL medium containing 20 ng/ml BMP4 (Novoprotein), 20 ng/ml activin A (Novoprotein) and 1.5 μM CHIP99021 (MCE) for 3 days (d) to allow differentiation into the mesoderm. hESC-CMs were differentiated from the mesoderm, differentiated in LI-APEL medium supplemented with XAV939 (MCE) for 3 d and then maintained in LI-APEL for another 3 d. From Day 12, DMEM (Gibco) supplemented with sodium-L-lactate was added, and hESC-CMs were harvested until Day 20 [ ]. hESC-ECs were also differentiated from the mesoderm.

Techniques: Immunofluorescence, Staining, Microscopy, Membrane, Fluorescence, In Vitro, CCK-8 Assay

Journal: Journal of Translational Medicine

Article Title: Interpenetrating network hydrogel-loaded embryonic stem cell-derived endocardial cells improves cardiac function after myocardial infarction

doi: 10.1186/s12967-025-06603-2

Figure Lengend Snippet: hESC-ECCs enhance the antioxidant stress of hESC-CMs and promote the angiogenesis and migration ability of hESC-ECs. A - C : Coculture for detecting the effect of hESC-ECCs on the antioxidant stress of hESC-CMs; A : hESC-CMs were cultured in LI-APEL medium until they reached 80% confluence and were divided into three groups: hESC-CMs cultured in LI-APEL medium as a control, hESC-CMs treated with H 2 O 2 , and hESC-CMs cocultured with hESC-ECCs for 48 h in transwell plates before being treated with H 2 O 2 ; B : Cck-8 test was performed on hESC-CMs after H 2 O 2 treatment, and the survival rates of hESC-CMs were measured by comparison with those of the control groups ( n = 6, ** P < 0.01), C : Images of hESC-CMs (×10) and TEM (×1200) of mitochondria in hESC-CMs; D - E : Coculture for detecting the effects of hESC-ECCs on the migration of hESC-ECs. D : hESC-ECs were suspended in medium and cultured in the cell plates until they reached 90% confluence, and a scratch was made on the bottom of the plate. Then, hESC-ECCs were cocultured with the hESC-ECs in transwell plates for 24 h and observed under a microscope (×4, 1, 3: Scratch image before coculture as a control; 2: hESC-ECs without coculture after 24 h of scratching; 4: hESC-ECs cocultured with hESC-ECCs after 24 h of scratching); E : Statistical analysis of the migration rate of hESC-ECs ( n = 8, * P ≤ 0.05). F : hESC-ECs were suspended in medium and cultured in the cell plates for 2 h, and then hESC-ECCs were cocultured with the hESC-ECs for 24 h and observed under a microscope (×4, 1: Control; 2: hESC-ECs cocultured with hESC-ECCs); G - J : Statistical analyses of the number of nodes, number of junctions, total branching length and total number of master segments of hESC-ECs ( n = 3). The data are shown as the means ± SDs, and differences among multiple groups were analyzed via one-way ANOVA. Differences between two groups were analyzed via Student’s t test

Article Snippet: For CM and EC differentiation, hESCs (Cellapy) were treated with LI-APEL medium containing 20 ng/ml BMP4 (Novoprotein), 20 ng/ml activin A (Novoprotein) and 1.5 μM CHIP99021 (MCE) for 3 days (d) to allow differentiation into the mesoderm. hESC-CMs were differentiated from the mesoderm, differentiated in LI-APEL medium supplemented with XAV939 (MCE) for 3 d and then maintained in LI-APEL for another 3 d. From Day 12, DMEM (Gibco) supplemented with sodium-L-lactate was added, and hESC-CMs were harvested until Day 20 [ ]. hESC-ECs were also differentiated from the mesoderm.

Techniques: Migration, Cell Culture, Control, CCK-8 Assay, Comparison, Microscopy