Fig. 1

A single nsPEF treatment (5 pulses, less than 10 s) can enhance the differentiation of MSCs. a Schematic of MSCs stimulated by nsPEFs. One million of MSCs were suspended in culture medium in gap cuvette and were subjected to 5 pulses of nsPEFs (i.e., 10 ns at 20 kV/cm and 100 ns at 10 kV/cm). And the time interval between two pulses is 1 s. Then, the trilineage differentiation induction was carried out. b Alizarin Red S, Oil red O staining, and Alcian blue staining for osteogenic, adipogenic, and chondrogenic differentiation at day 14, insets show the no-staining counterparts. c Quantification of differentiation into osteogenic, adipogenic, and chondrogenic lineages. (3 batches of studies were tested with 3 biological donors, values are mean ± SEM from one representative batch with 5 technical repeats, one-way ANOVA, *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, NS, p > 0.05) d–f qRT-PCR for the mRNA levels of genes associated with trilineage differentiation (osteogenic: RUNX2, OCN; adipogenic: PPARγ, LPL; chondrogenic: SOX9, COLII) respectively at day 14. (3 batches of studies were tested with 3 biological donors, values are mean ± SEM from one representative batch with 5 technical repeats, one-way ANOVA, *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, NS, p > 0.05)