Purpose <p>Human adipose tissue-derived stem cells (hADSCs) have emerged as a promising source of cells for tissue engineering and regenerative medicine. However, their differentiation potential is restricted and requires enhancements. This study explores the reprogramming of hADSCs through exogenous induction of the miR-302/367 cluster.</p> Methods <p>Human ADSCs were transfected with the mock or miR-302/367 cluster-expressing vectors. One week after transfection, expression levels of several pluripotency-related genes, epithelial-to-mesenchymal (EMT) markers, and mechanistic target of rapamycin kinase (mTOR) signaling factors were assessed by qPCR and western blot. Additionally, the influence of miR-302/367 cluster overexpression on the proliferation and adipogenic differentiation of the ADSCs was evaluated.</p> Results <p>One week after transfection, the expression of several pluripotency-related genes and epithelial markers was significantly upregulated, while mesenchymal markers were downregulated in the miR-302/367-transfected cells compared with the mock group. Additionally, the levels of several mTOR signaling factors were reduced in the miR-302/367-transfected ADSCs. Flow cytometry analysis showed a decrease in the abundance of ADSCs in the S phase and an increase in the population of cells in the G1 phase of the cell cycle. Moreover, the adipogenic differentiation of the miR-302/367-transfected cells was significantly enhanced.</p> Conclusion <p>The overexpression of the miR-302/367 cluster directed the ADSCs towards a more pluripotent state and promoted their adipogenic potential. However, miR-302/367 overexpression diminished the proliferative capacity of hADSCs, which warrants a comprehensive investigation. Further evaluations are needed to fully elucidate the differentiation potential and regenerative capacity of the ADSCs reprogrammed by the miR-302/367 cluster before any clinical application.</p>