| US 7,572,631 B2 | ||
| Activation and expansion of T cells | ||
| Ronald J Berenson, Mercer Island, Wash. (US); Che Law, Shoreline, Wash. (US); Mark Bonyhadi, Issaquah, Wash. (US); Zahbea Saund, Seattle, Wash. (US); Stewart Craig, Issaquah, Wash. (US); Alan Hardwick, Seattle, Wash. (US); Dale Kalamasz, Redmond, Wash. (US); David McMillen, Seattle, Wash. (US); and Harjinder Singh Chana, Issaquah, Wash. (US) | ||
| Assigned to Invitrogen Corporation, Carlsbad, Calif. (US) | ||
| Filed on Mar. 02, 2005, as Appl. No. 11/70,841. | ||
| Application 11/070841 is a continuation in part of application No. 10/762210, filed on Jan. 20, 2004. | ||
| Application 10/762210 is a continuation in part of application No. 10/350305, filed on Jan. 22, 2003, abandoned. | ||
| Application 10/350305 is a continuation in part of application No. 10/187467, filed on Jun. 28, 2002, abandoned. | ||
| Application 10/187467 is a continuation in part of application No. 10/133236, filed on Apr. 26, 2002, granted, now 6,867,041. | ||
| Application 10/133236 is a continuation in part of application No. 09/960264, filed on Sep. 20, 2001, granted, now 6,797,514. | ||
| Application 09/960264 is a continuation in part of application No. 09/794230, filed on Feb. 26, 2001, granted, now 6,905,874. | ||
| Claims priority of provisional application 60/184788, filed on Feb. 24, 2000. | ||
| Claims priority of provisional application 60/249902, filed on Nov. 17, 2000. | ||
| Prior Publication US 2006/0121005 A1, Jun. 08, 2006 | ||
| Int. Cl. C12N 5/00 (2006.01) | ||
| U.S. Cl. 435—375 [435/325; 435/372.3] | 7 Claims |
| 1. A method for expanding a population of T cells from a B-cell chronic lymphocytic leukemia (B-CLL) patient by cell surface
moiety ligation, comprising:
a. selecting a patient having B-CLL;
b. providing a population of cells from the patient wherein at least a portion thereof comprises T cells;
c. contacting said population of cells for about two hours with a surface, wherein said surface is a surface of a paramagnetic
particle and wherein said surface has attached thereto a first agent that ligates a TCR/CD3 complex of a T cell in said population
of cells, and a second agent that ligates a CD28 accessory molecule of said T cell;
d. applying a magnetic force that predominantly drives simultaneous T cell concentration and ligation of the TCR/CD3 complex
and the CD28 accessory molecule of said T cells;
e. seeding said population of cells at a concentration of between about 0.2×106 and 5.0 ×106 cells/ml in a closed system comprising a disposable bioreactor bag comprising at least one inlet filter and one outlet filter
and further contacting said T cells with said surface;
f. perfusing medium through said closed system at about 1 ml/minute staffing at about day 8;
g. rocking said bioreactor bag on a rocking platform at about 5-15 rocks/minute;
and wherein said T cells expand to a concentration of about between 15 ×106 cells/ml to about 50×106 cells/ml in less than about two weeks.
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