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US 2001/0055808 A1Published application
Use of collagenase in the preparation of neural stem cell cultures
OwnerNobuko Uchidainventor
Life of this patent
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Abstract
The invention provides a method for using collagenase to dissociate neural stem cells in neural stem cell cultures. The collagenase treatment results in an increased cell viability and an increased number of proliferated neural stem cells over time.
Claims37
A method for the in vitro proliferation of a neural stem cell culture comprising the steps of:
- (a) obtaining dissociated neural tissue in cell suspension, the suspension containing one or more multipotent neural stem cells capable of producing progeny that are capable of differentiating into neurons and glia;
- (b) culturing the cell suspension in a culture medium containing at least one proliferation-inducing growth factor to proliferate the neural stem cells in (a) to generate a neural stem cell culture;
- and
- (c) passaging the cell culture in (b) by treating the culture with an amount of a collagenase preparation effective to dissociate the cells in the culture and passing the cell culture to a culture medium containing at least one proliferation-inducing growth factor to further proliferate the neural stem cell culture.
1 more element
A method for the in vitro proliferation of a neural stem cell culture comprising the steps of:
- (a) obtaining dissociated neural tissue in cell suspension, the suspension containing one or more multipotent neural stem cells capable of producing progeny that are capable of differentiating into neurons and glia;
- (b) culturing the cell suspension in a culture medium containing at least one proliferation-inducing growth factor to proliferate the neural stem cells in (a) to generate a neural stem cell culture;
- and
- (c) passaging the cell culture in (b) by treating the culture with an amount of a collagenase preparation effective to dissociate the cells in the culture and passing the cell culture to a culture medium containing at least one proliferation-inducing growth factor to further proliferate the neural stem cell culture, wherein the percent viability of the cells in the culture is at least 60% after being passaged.
1 more element
A method for expanding a neural stem cell culture comprising the steps of:
- (a) culturing a neural stem cell culture that contains one or more multipotent neural stem cells capable of producing progeny that are capable of differentiating into neurons and glia in a culture medium containing at least one proliferation-inducing growth factor to proliferate the culture;
- and
- (b) passaging the cell culture in (a) by treating the culture with an amount of a collagenase preparation effective to dissociate the cells in the culture and passing the cell culture to a culture medium containing at least one proliferation-inducing growth factor to further proliferate the neural stem cell culture, wherein there is at least a three-fold increase in cell number between weekly passages.
A method for expanding a neural stem cell culture comprising the steps of:
- (a) culturing a neural stem cell culture that contains one or more multipotent neural stem cells capable of producing progeny that are capable of differentiating into neurons and glia in a culture medium containing at least one proliferation-inducing growth factor to proliferate the culture;
- and
- (b) passaging the cell culture in (a) by treating the culture with an amount of a collagenase preparation effective to dissociate the cells in the culture and passing the cell culture to a culture medium containing at least one proliferation-inducing growth factor to further proliferate the neural stem cell culture, wherein the accumulated cell number can be expanded from 106 cells to at least 109 cells in less than 49 days.
Dependencies are inferred from the claim text.