Mesenchymal stromal cells (MSCs) play a critical role in supporting hematopoietic stem and progenitor cells (HSPCs) within the bone marrow (BM) niche by providing a specialized extracellular matrix and essential signaling cues. Through the secretion of key ECM proteins and cell adhesion molecules, MSCs regulate HSPC proliferation, differentiation, homing, retention, and maintain their quiescence necessary for effective hematopoiesis. Understanding the intricate dynamics between MSCs and HSPCs in the BM is essential for advancing HSPC-based therapies. Targeting niche components, such as ECM proteins and CAMs, can enable HSPC expansion and engraftment, benefiting hematopoietic stem cell transplantation and gene therapy outcomes. In this review, we provide an overview of the niche components involved in MSCs-HSPCs interactions and its co-culture in improving the therapeutic outcomes, discuss existing challenges and propose possible solutions.