The lining epithelium of the stomach includes multipotent stem cells which undergo proliferation and migration-associated differentiation. These cells give rise to multiple cell lineages that produce mucus, acid, pepsinogen and various hormones/peptides. A 3D culture for stem cells would facilitate identification of the factors that control proliferation and/or differentiation programs. Here, we report on the use of disk-like ImmobaSil HD silicone-rubber matrix based microcarriers that are permeable to oxygen and reduce the creation of toxic environment within the center of the microcarrierd for culturing the mouse gastric stem (mGS) cells. We define several parameters that affect the initial cell attachment such as size of cell inoculum, serum concentration, mode and speed of agitation. We show that although such a microcarrier allows for attachment and growth of gastric stem cells, it does not lend itself and does not support the functional differentiation of such cells.