Methods for preparing therapeutically active cells using microfluidics
The present invention is directed to the use of microfluidics in the preparation of cells and compositions for therapeutic uses.
1. A method for preparing a purified leukocyte composition comprising:
a) purifying leukocytes from an apheresis sample, wherein the sample comprises leukocytes and platelets and the leukocytes are separated from the platelets by:
i) performing a size-based separation using a microfluidic device configured to separate cells by deterministic lateral displacement to produce an enriched product in which, compared to the sample, the percentage of platelets has decreased and the percentage of leukocytes has increased;
ii) in addition to the size-based separation, performing an affinity-based separation by binding one or more types of leukocytes to a carrier that binds to the one or more types of leukocytes with specificity, and then separating the carrier-bound cells from cells not bound to carrier;
b) after the purification of step a), activating and expanding the purified leukocytes in culture to produce the purified leukocyte composition.
2. The method of claim 1 , wherein the apheresis sample is obtained from a patient and the patient has rheumatoid arthritis, type I diabetes, or multiple sclerosis.
3. The method of claim 1 , wherein, after the purified leukocyte composition is prepared, said composition is administered to a patient.
4. The method of claim 1 , wherein the platelets in the enriched product of paragraph a)i) are depleted by at least 80% compared to the sample and/or there are no more than 5 platelets per leukocyte in the enriched product.
5. The method of claim 1 , further comprising:
c) genetically engineering the activated leukocytes.
6. The method of claim 1 , wherein the apheresis sample is obtained from a patient and the patient has cancer.
7. The method of claim 1 , wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: B-cells, NK-cells, and monocytes.
8. The method of claim 1 , wherein the leukocytes in the purified leukocyte composition are granulocytes.
9. The method of claim 1 , wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: central memory T cells; helper T cells and regulatory T cells.
10. The method of claim 1 , wherein the sample was obtained by leukapheresis.
11. The method of claim 1 , wherein the cells of the purified leukocyte composition are transferred into a pharmaceutical composition and are not frozen prior to transfer.
12. The method of claim 5 , wherein the apheresis sample is obtained from a patient and the patient has cancer.
13. The method of claim 12 , wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: B-cells, NK-cells, and monocytes.
14. The method of claim 12 , wherein the leukocytes in the purified leukocyte composition are granulocytes.
15. The method of claim 12 , wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: central memory T cells; helper T cells and regulatory T cells.
16. The method of claim 12 , wherein the sample was obtained by leukapheresis.
17. The method of claim 12 , wherein the cells of the purified leukocyte composition are transferred into a pharmaceutical composition and are not frozen prior to transfer.