A method of preparing a stromal cell conditioned medium useful in
expanding undifferentiated hemopoietic stem cells to increase the number
of the hemopoietic stem cells is provided. The method comprising: (a)
establishing a stromal cell culture in a stationary phase plug-flow
bioreactor under continuous flow on a substrate in the form of a sheet,
the substrate including a non-woven fibrous matrix forming a
physiologically acceptable three-dimensional network of fibers, thereby
expanding undifferentiated hemopoietic stem cells; and (b) when a desired
stromal cell density has been achieved, collecting medium from the
stationary phase plug-flow bioreactor, thereby obtaining the stromal cell
conditioned medium useful in expanding the undifferentiated hemopoietic
stem cells.