Culturing human bone marrow stromal cells

Bone marrow (BM) stromal cells (BMSC) comprise hematopoietic and non-hematopoietic cells, such as macrophages, fibroblasts, adipocytes, and endothelial cells. These cells are considered to be hematopoietic supporting cells and have been used to assess functions within the BM microenvironment using in vitro models. These cells are critical when evaluating the functions of hematopoietic stem cells in vitro using the long-term culture initiating assay. Unlike solid organs that can be easily removed for research purposes, the BM cannot be similarly harvested. Thus, to understand bone marrow functions, model systems require the hematopoietic stem and progenitor cells to be studied within the endogenous microenvironment. The collection of cells and factors within the stromal culture represent this niche environment. In this chapter, we present a step-by-step protocol for isolating stromal cultures from bone marrow aspirates.