<p>Exosomes derived from plant cell suspension cultures are a promising resource for biomolecule transport and therapeutic applications. This study aimed to increase the yield of exosomes from loquat suspension cells (LSCs) and characterize their microRNAs (miRNAs), laying a foundation for future applications. The LSCs were sampled at the late logarithmic, early plateau, and late plateau phases. The LSCs were treated with 15–35°C for 72&#xa0;h in the late plateau phase; control LSCs were maintained at 25°C. LSC-derived exosomes were isolated and their morphology, average particle size, and number were compared. High-throughput sequencing was conducted to analyze the miRNAs in exosomes isolated from LSCs and loquat leaves. Quantitative PCR was employed to examine the expression profiles of selected miRNAs in different samples, growth phases, and temperature treatments. No significant differences in exosome morphology and average particle size were detected among the treatments. Treatment at 35°C for 72&#xa0;h significantly increased the exosome particle number to more than three times that of the control. High-throughput sequencing identified 45 differentially expressed miRNAs from 13 families, corresponding to 60 target genes. The expression patterns of seven randomly selected miRNAs were consistent with the initial miRNA-sequencing samples. The expression levels of miR-390, miR-397, and miR-408 in LSC-derived exosomes exhibited distinct dynamic changes among growth phases and temperature treatments. This study provides insights into a method for increasing the particle number of exosomes from LSCs, and characterizes their miRNA composition and expression profiles, thereby contributing to innovation in exosome production from plant suspension cultures.</p>