Tangential flow filtration devices and methods for leukocyte enrichment
First Claim
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1. A method for preparing monocytic dendritic cell precursors from a sample of blood constituents from a subject wherein the sample comprises leukocytes, the method comprising:
- (1) introducing the sample into a remover unit, wherein the remover unit comprises a cross-flow chamber, a filtrate chamber, and a filter disposed therebetween;
the filter having a retentate surface and a filtrate surface, the filter in fluid communication with the cross-flow chamber and the filtrate chamber;
the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising the sample of blood constituents comprising the leukocytes from a source into the cross-flow chamber and parallel to the retentate surface of the filter, and the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;
(2) subjecting the sample to cross-flow substantially parallel to a filter having a pore size of about 1 to about 10 microns;
(3) subjecting the sample to filtration through the filter;
(4) selectively removing non-leukocyte blood constituents from the sample through the filter and removing the leukocytes through the centrally disposed outlet to form a cell population enriched for leukocytes; and
(5) preparing monocytic dendritic cell precursors from the cell population enriched for leukocytes.
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Abstract
The present invention provides tangential flow filtration devices and methods for enriching a heterogenous mixture of blood constituents for leukocytes by removal of non-leukocyte blood constituents. In one particular embodiment the device can provide a composition enriched in monocytes.
42 Citations
24 Claims
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1. A method for preparing monocytic dendritic cell precursors from a sample of blood constituents from a subject wherein the sample comprises leukocytes, the method comprising:
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(1) introducing the sample into a remover unit, wherein the remover unit comprises a cross-flow chamber, a filtrate chamber, and a filter disposed therebetween;
the filter having a retentate surface and a filtrate surface, the filter in fluid communication with the cross-flow chamber and the filtrate chamber;
the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising the sample of blood constituents comprising the leukocytes from a source into the cross-flow chamber and parallel to the retentate surface of the filter, and the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;(2) subjecting the sample to cross-flow substantially parallel to a filter having a pore size of about 1 to about 10 microns; (3) subjecting the sample to filtration through the filter; (4) selectively removing non-leukocyte blood constituents from the sample through the filter and removing the leukocytes through the centrally disposed outlet to form a cell population enriched for leukocytes; and (5) preparing monocytic dendritic cell precursors from the cell population enriched for leukocytes. - View Dependent Claims (2, 3, 4, 5)
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6. A method for enriching a sample of blood constituents for dendritic cells, comprising:
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(1) introducing the sample into a tangential flow filtration (TFF) unit, the TFF unit comprising a cross-flow chamber, a filtrate chamber, and a filter in fluid communication with the cross-flow chamber and the filtrate chamber, the filter having a retentate surface and a filtrate surface, the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising the sample of blood constituents comprising leukocytes from a source into the cross-flow chamber and parallel to the retentate surface of the filter, and the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface, the filter having a pore size of about 1 to about 10 microns; (2) recirculating the sample through the TFF unit at a predetermined input rate and a predetermined filtration rate, the predetermined input rate at least five times the predetermined filtration rate; (3) isolating a cell population enriched for leukocytes; and (4) preparing dendritic cells from the enriched leukocyte cell population. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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Specification