Résumé : Stem cell therapy is a potential method for the treatment of numerous diseases. The most frequent cellular source is bone marrow-derived mesenchymal stromal cells (BM-MSC). Human adipose-derived stromal cells (ADSC) share similar properties with BM-MSC as they support hematopoiesis, modulate ongoing immune responses and differentiate into cells of mesodermal origin. On the other hand, ADSC have higher frequency in situ, higher availability and very few ethical issues compared to BM-MSC giving them an advantage over BM-MSC for clinical use. Most of the methods used to isolate ADSC contain a collagenase digestion step, but the type of collagenase and time of sample digestion vary among studies and these differences could have an impact on the cell properties and thus in result comparison. To overcome this obstacle, we propose a new method to isolate ADSC from lipoaspirate without collagenase digestion step. We compared ADSC obtained with our method versus classical protocol using collagenase digestion. Cells obtained with our method are equivalent but has a better long term hematopoietic support than those obtained with classical method. Moreover, our method has an advantage over the classical one as it is easier, safer, faster, less expensive and more consistent with good manufacturing practices (GMP) to obtain large number of ADSC ex vivo.