2003 Fifth Annual Beckman Scholars Symposium
Arnold and Mabel Beckman Foundation

The Effect of Nerve Growth Factor on the Differentiation of Muscle Derived Stem Cells

Nima Salari
University of Pittsburgh

We have investigated whether the extent of pluripotential differentiation of muscle-derived stem cells (MDSC, derived by the pre-plate technique from the tissue of adult mice) can be controlled in vitro by use of growth factors. The influence of the nerve growth factor (NGF) was investigated when it was used in different doses (0, 50, and 100 ng/ml) to stimulate the cells of three different populations at varying passages (FN14, FT27, MN44 at passages 14, 27, and 44 respectively) for 7 days. The expression of the stem cell neurogenic markers was assayed to determine the degree of the influence of NGF on MDSC. The NGF-treated cells were assayed by immunocytochemistry for the expression of specific markers for Schwann cell (CNPase), and neurons (NeuN, ß-Tubulin). The percentage of positive cells was determined as the ratio of positive cells to total number of cells in 5 randomly chosen fields at 200x magnification.

Interestingly, a low percentage of CNPase positive cells were detected in all cultures without NGF stimulation. However, after stimulation with NGF, CNPase positive cells increased in 2 of the 3 populations (FN14 and FT27), however the third population (MN44) showed no apparent responsiveness to the stimulation. It is also interesting to mention that the population of FT27 showed a high variability in responsiveness to the stimulation where the range of data reached from a low responsiveness to a fairly high responsiveness. Similar results were obtained when the expression of NeuN and ß-Tubulin was analyzed by immunocytochemistry.

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