Method of producing undifferentiated hemopoietic stem cells using a stationary phase plug-flow bioreactor

Patent Number: 6911201

One of the limiting factors to clinical use of autologous hematopoietic stem cells, as well as cord blood cells, is the limited ways of expanding them. This patent addresses this issue by providing a novel system for such expansion that appears to be more "in vivo-like". Essentially, the patent has 2 independent claims covering:


A method of expanding undifferentiated hemopoietic stem cells to increase the number of the hemopoietic stem cells, the method comprising the steps of:

(a) culturing in a stationary phase plug-flow bioreactor a stromal cell culture under continuous flow of a culture medium on a substrate in the form of a sheet, said substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers to thereby generate a three dimensional stromal cell culture; and

(b) seeding undifferentiated hemopoietic stem cells into said stationary phase plug-flow bioreactor including said three dimensional stromal cell culture and under said continuous flow of said culture medium, thereby expanding the undifferentiated hemopoietic stem cells to increase the number of the hemopoietic stem cells without the hemopoietic stem cells undergoing differentiation to obtain expanded undifferentiated hemopoietic stem cells.

A method of transplanting undifferentiated hemopoietic stem cells into a recipient, the method comprising the steps of: (a) expanding the undifferentiated hemopoietic stem cells to increase the number of the hemopoietic stem cells by: (i) culturing in a stationary phase plug-flow bioreactor a stromal cell culture under continuous flow of a culture medium on a substrate in the form of a sheet, said substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers to thereby generate a three dimensional stromal cell culture; and (ii) seeding undifferentiated hemopoietic stem cells into said stationary phase plug-flow bioreactor including said three dimensional stromal cell culture and under a continuous flow of a culture medium, thereby expanding the undifferentiated hemopoietic stem cells increase the number of the hemopoietic stem cells without the hemopoietic stem cells undergoing differentiation to obtain expanded undifferentiated hemopoietic stem cells; and (b) transplanting the undifferentiated hemopoietic stem cells resulting from step (a) in the recipient.

View this patent on the USPTO website.