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Human lineage committed cell composition with enhanced proliferative potential, biological effector function, or both: methods for obtaining same; and their usesRelated Patent Categories: Drug, Bio-affecting And Body Treating Compositions, Whole Live Micro-organism, Cell, Or Virus ContainingHuman lineage committed cell composition with enhanced proliferative potential, biological effector function, or both: methods for obtaining same; and their uses description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20060093581, Human lineage committed cell composition with enhanced proliferative potential, biological effector function, or both: methods for obtaining same; and their uses. Brief Patent Description - Full Patent Description - Patent Application Claims
BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention relates to methods for culturing lineage committed human cells, the cells-thus obtained, and their uses. [0003] 2. Description of the Background [0004] There is significant interest using both early and lineage committed cells for a variety of therapeutic purposes. [0005] In tissue engineering, the goal is to reconstitute fully or partially functioning human tissue in vitro to enable a variety of clinical and other applications. Several studies have been carried out that are aimed at reconstituting functioning human tissues in vitro. Illustratively, the cultivation of human skin has been successful. [0006] The hematopoietic system exemplifies the broad range of cells involved in protection of mammalian hosts against pathogens toxins, neoplastic cells, and other diseases. The hematopoietic system is believed to evolve from a single stem cell, from which all the lineages of the hematopoietic system derive. Hematopoietic cells have been used in human therapy. Methods and apperati for culturing precursor hematopoietic cells to obtain desired mature hematopoietic cells have been described. See, U.S. Pat. Nos. 5,605,822, 5,399,493; 5,437,994; 5,459,069; 5,635,386, 5,670,147 and 5,670,351. [0007] Adoptive cell therapy is the ex vivo expansion and re-infusion of immune effector cells into human recipients for the treatment or prevention of disease (Rosenberg et al., Science 233, 1318 (1986)). Developments in T-lymphocyte-based adoptive immunotherapy have lead to significant advances in the treatment of metastatic cancer (Rosenberg et al., Science 233, 1318 (1986); Rosenberg et al., N. Eng. J. Med. 319, 1676 (1988); Rosenberg et al., N. Eng. J. Med. 323, (1990)) and viral diseases including Epstein-Barr Virus (EBV) (Heslop et al. 1996), cytomegalovirus (CMV) (Riddell et al., Science 257, 238 (1992)) and human immunodeficiency virus (HIV) (Whiteside et al., Blood 81, 2085 (1993); Ridell et al., Hum. Gen. Ther. 3, 319 (1992)) in humans. Multiple patient populations have been identified in which T-cell therapy has achieved a response rate of approximately 34% after administration of tumor infiltrating lymphocytes (TILs) to metastatic melanoma patients (Rosenberg et al., J. Natl. Cancer Inst. 86, 1159 (1994)). A target human therapeutic dose of 10.sup.10-10.sup.11 TILs is suggested based on extrapolation from studies in mouse tumor models and clinical experience (Topalian et al., J. Immunol. Meth. 102, 107 (1987)). Clinical responses to melanoma tumors have been obtained after expansion ex vivo of TILs in medium containing interleukin-2 (IL-2) and re-infusion of greater than 10.sup.11 T-cells per treatment cycle together with high doses of exogenous IL-2 (Rosenberg et al., N. Eng. J. Med. 319, 1676 (1988); Rosenberg et al. 1994). The enhanced anti-tumor activity of TILs in vivo compared to lymphokine activated killer (LAK) cells or NK cells may be a function of several complex processes including the ability of these T-cells to proliferate and release an array of lymphokines, to recirculate and accumulate at sites of tumor growth, and to specifically recognize and lyse autologous tumor cells (Pockaj et al. 1994; Rosenberg et al. 1994). [0008] Dendritic cells (DCs) are highly specialized bone-marrow derived antigen presenting cells (APCs) which function as potent stimulating cells for primary T-lymphocyte-mediated immune response. Dendritic cell based immunotherapy is an emerging treatment strategy involving ex vivo expansion and reinfusion of antigen-pulsed or genetically modified DCs into human patients to vaccinate against cancer or infectious diseases. Clinical implementation of these therapies requires the capability of product sufficient quantities of functional DCs for effective patient treatment. [0009] The use of cultured human cells in human therapy has required that a quantity of active cells sufficient to provide a therapeutic effect upon infusion into the patient be used. This has required using culture systems providing large numbers of cells. There is therefore a need for methods for obtaining lineage committed cells with augmented proliferative potential, biological function, or both since such methods would provide more potent cell compositions, capable of being used in smaller amounts in therapy. SUMMARY OF THE INVENTION [0010] Accordingly, it is an object of this invention to provide novel methods, including culture media conditions, for obtaining human lineage committed cells with enhanced proliferative potential, biological function, or both. [0011] It is another object of this invention to provide cultured human lineage committed cells having enhanced proliferative potential, biological function, or both. [0012] These objects and others may be accomplished by a method for obtaining lineage committed human cells having enhanced proliferative potential, biological function, or both. In this method, lineage committed human cells are cultured in any physiologically acceptable liquid culture medium conditions with the liquid culture medium being replaced. BRIEF DESCRIPTION OF THE FIGURES [0013] FIG. 1 shows the amount of cytokine produced after stimulation of T-cells cultured using continuous medium exchange conditions according to the present invention and cells cultured under standard static low density medium conditions. AIM V represents a control with no added stimulus, all experiments were cultured with AIM V culture medium. OKT3 represents anti-CD3 mAb. (A) IFN-.gamma. production; (B) TNF-.alpha. production; (C) GM-CSF production; (D) IL-10 production. [0014] FIG. 2 shows (A) the lactate concentration during a dendritic cell culture according to the present invention (100% exchange) and in a static culture (0% medium exchange) at low inoculum density, and (B) the mixed leucocyte response (MLR) of the isolated dendritic cells. [0015] FIG. 3 shows (A) the lactate concentration during a dendritic cell culture according to the present invention (150% exchange) and in a static culture (0% medium exchange) at intermediate inoculum density, and (B) the mixed leucocyte response (MLR) of the isolated dendritic cells. [0016] FIG. 4 shows (A) the lactate concentration during a dendritic cell culture according to the present invention at 100% exchange and 300% medium exchange at high inoculum density, and (B) the mixed leucocyte response (MLR) of the isolated dendritic cells. [0017] FIG. 5 shows the effect of medium exchange rate of canine chondrocyte production at different inoculum densitites. DESCRIPTION OF THE PREFERRED EMBODIMENTS [0018] The present invention applies to the culture of lineage committed human cells using any known culture techniques. The invention is based on the discovery that by replacing the liquid culture media in any lineage committed human cell culture without substantially changing the cell density, it is possible to obtain cells having an enhanced proliferative potential, an enhanced biological function, or both. The cells obtained in accordance with the invention have enhanced proliferative potential as compared to the cells prior to culture and/or as compared to cells one obtains using otherwise identical culture conditions except that the liquid culture media is replaced by diluting the culture to achieve a lower cell density. Similarly, the biological function of the cells obtained in accordance with the invention is enhanced as compared with the biological function of these same cells prior to culture and/or as compared to the biological function of the same type of cells cultured under otherwise identical culture conditions except that the liquid medium is not replaced. [0019] The cells one obtains in accordance with the invention are, by virtue of their enhanced proliferative potential and/or biological effector function, more potent. The present invention permits using a smaller number of lineage committed human cells to obtain similar or better results than one obtains using lineage committed human cells cultured in accordance with techniques existing prior to the present invention. The lineage committed human cells one obtains in accordance with the invention can be used in any application in which lineage committed human cells are used, including, but not limited to, therapy treatments including adoptive immunotherapy with effector cells, tumor specific cytotoxic T-cells, infectious disease specific cytotoxic T lymphocytes, cytokine induced killer cell therapy, antigen presenting cells to either tumor or infectious diseases, dendritic cells, antigen primed, dendritic cells, tumor vaccines, genetically modified attenuated tumor cells, genetically modified antigen presenting cells, structural repair procedures such as cartilage defect repair, bone defect repair, tissue repair, would healing, burn care, solid organ repair, neurological defect repair, etc. Derivation and expansion of antigen specific T-cell populations including, but not limited to, viral (e.g. EBV, HPV, CMV, HIV, influenza) and tumor reactive T-cells, ex vivo expansion of human tumor infiltrating lymphocytes (TILs) for adoptice cancer immunotherapy, derivation and ex vivo expansion of cytokine induced killer (CIK) cells for rejection of human tumors including leukemia, lymphoma, breast and other cancers. Also, immunotherapy after autologous or allogeneic bone marrow transplantation. With dendritic cells: antigen presenting cell (APC)-based vaccines to stimulate T-cell responses in vitro or in vivo against simple or complex antigens including, but not limited to, tumor, viral, fungal and bacterial antigens. Therapy may include treatment or prevention of disease in normal individuals or human cancer patients. Lineage Committed Cells Continue reading about Human lineage committed cell composition with enhanced proliferative potential, biological effector function, or both: methods for obtaining same; and their uses... 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