Methods for Enhanced Propagation of Cells
First Claim
1. A method for enhanced propagation of cells from a kidney sample, the method comprising promoting expansion of renal precursor cells from the sample, thereby to enhance propagation of a cell population from the sample.
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Abstract
The present invention relates generally to methods for the isolation and propagation of cells. For example, embodiments of the present invention relate to isolation and propagation methods for the manufacture of a large number of cells for use, for example, in biotherapeutic devices, such as devices for renal replacement therapy for the treatment of acute renal failure (ARF), acute tubular necrosis (ATN), multi-organ failure (MOF), sepsis, cardiorenal syndrome (CRS) and end-stage renal disease (ESRD).
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Citations
21 Claims
- 1. A method for enhanced propagation of cells from a kidney sample, the method comprising promoting expansion of renal precursor cells from the sample, thereby to enhance propagation of a cell population from the sample.
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10. A method for propagating renal cells from a kidney sample, the method comprising:
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(a) treating the kidney sample to produce tissue fragments having a dimension of about 212 μ
m or less in size;(b) harvesting at least a portion of the tissue fragments by centrifugation at greater than about 50×
(g) to produce a pelleted slurry;(c) applying a portion of the pelleted slurry produced in step (b) to a solid support and culturing the cells from the pelleted slurry in the absence of retinoic acid and under conditions to permit the renal cells to attach to the solid support and divide; (d) harvesting the cells produced in step (c) prior to reaching confluence on the solid support; (e) applying a portion of the cells harvested in step (d) to a solid support and subculturing the cells in the absence of retinoic acid and under conditions to permit the renal cells to attach to the solid support and divide; and (f) harvesting the cells produced in step (e) to produce a propagated population of renal cells. - View Dependent Claims (11, 18, 19, 20, 21)
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12. A method for enhanced propagation of cells from a sample, the method comprising:
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separating the sample into fragments of about 212 μ
m or less in size to produce a slurry;centrifuging the slurry at greater than about 50×
(g) to produce a pelleted slurry;creating a culture by applying to a growth surface about 10 μ
l or less of the pelleted slurry per square centimeter of surface area on which the culture is to be grown; andsubculturing about 8.7×
104 or fewer cells from the culture per square centimeter of surface area on which the subculture is to be grown. - View Dependent Claims (13, 14, 15, 16, 17)
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Specification