Effects of fibroblasts conditioned media on differentiation of programmable cells of monocytic origin to insulin-producing cells

ABSTRACT

The characteristic of stem cells in self renewal and differentiation to different types of cells has stimulated the interests for using stem cells as a starting material for generating insulin secreting cells. We've evaluated the differentiation potential of Programmable cells of monocytic origin [PCMOs] into insulin producing cells effected from the growth factors and fibroblasts conditioned media [FCM]. Peripheral blood monocytes of rat were cultured for 6 days in RPMI with 15% FBS, beta- mercaptoethanol, MCSF and interleukin-3. Then, these cells were incubated in differentiation media with HGF, EGF, Nicotinamide, 15% fibroblasts conditioned media and glucose for 15days. Morphological differences of cells were studied by invert microscope. In several stages, the amounts of insulin in supernatant of cells were measured by radioimmunoassay kit. Also productions of insulin from differentiated cells were studied with DTZ special staining. In response to MCSF and IL-3, monocytes dedifferentiated. These programmable cells of monocytic origin [PCMOs] were capable of differentiating into insulin producing cells in differentiation media. The morphology of differentiated cells was similar to Beta cells and the amount of insulin in supernatant of differentiated cells was much higher than PCMOs [P<0.05]. HGF, EGF, Nicotinamide and fibroblasts conditioned media are differentiation factors of PCMOs into insulin producing cells. According to the results insulin producing cells can be differentiated from programmable cells of monocytic origin in presence of fibroblasts conditioned media