Methods of growing tumor infiltrating lymphocytes in gas-permeable containers

Abstract

An embodiment of the invention provides a method of promoting regression of cancer in a mammal comprising obtaining a tumor tissue sample from the mammal; culturing the tumor tissue sample in a first gas permeable container containing cell medium therein; obtaining tumor infiltrating lymphocytes (TIL) from the tumor tissue sample; expanding the number of TIL in a second gas permeable container containing cell medium therein using irradiated allogeneic feeder cells and/or irradiated autologous feeder cells; and administering the expanded number of TIL to the mammal. Methods of obtaining an expanded number of TIL from a mammal for adoptive cell immunotherapy are also provided.

1 . A method of obtaining an expanded number of TIL from a mammal for adoptive cell immunotherapy comprising:
 obtaining a tumor tissue sample from the mammal;   culturing the tumor tissue sample in a first gas permeable container containing cell medium therein;   obtaining tumor infiltrating lymphocytes (TIL) from the tumor tissue sample;   expanding the number of TIL in a second gas permeable container containing cell medium therein using irradiated allogeneic feeder cells and/or irradiated autologous feeder cells.   
     
     
         2 . A method of promoting regression of cancer in a mammal comprising:
 obtaining a tumor tissue sample from the mammal;   culturing the tumor tissue sample in a first gas permeable container containing cell medium therein;   obtaining TIL from the tumor tissue sample;   expanding the number of TIL in a second gas permeable container containing cell medium therein using irradiated allogeneic feeder cells and/or irradiated autologous feeder cells; and   administering the expanded number of TIL to the mammal.   
     
     
         3 . A method of obtaining an expanded number of TIL from a mammal for adoptive cell immunotherapy comprising:
 obtaining a tumor tissue sample from the mammal;   obtaining TIL from the tumor tissue sample;   expanding the number of TIL in a gas permeable container containing cell medium therein using irradiated allogeneic feeder cells and/or irradiated autologous feeder cells.   
     
     
         4 . A method of promoting regression of cancer in a mammal comprising:
 obtaining a tumor tissue sample from the mammal;   obtaining TIL from the tumor tissue sample;   expanding the number of TIL in a gas permeable container containing cell medium therein using irradiated allogeneic feeder cells and/or irradiated autologous feeder cells; and   administering the expanded number of TIL to the mammal.   
     
     
         5 . The method of  claim 3 , further comprising selecting TIL capable of lysing cancer cells. 
     
     
         6 . The method of  claim 1 , wherein culturing the tumor tissue sample comprises culturing the tumor tissue sample submerged under a height of at least about 1.3 cm of cell culture medium in the first gas permeable container. 
     
     
         7 . The method of  claim 1 , wherein culturing the tumor tissue sample comprises culturing the tumor tissue sample submerged under a height of at least about 2.0 cm of cell culture medium in the first gas permeable container. 
     
     
         8 . The method of  claim 1 , wherein culturing the tumor tissue sample comprises culturing the tumor tissue sample in a gas permeable container having at least about 650 cm 2  of cell culture surface area. 
     
     
         9 . The method of  claim 1 , wherein the cancer is melanoma. 
     
     
         10 . The method of  claim 1 , wherein expanding the number of TIL comprises culturing the TIL submerged under a height of at least about 1.3 cm of cell culture medium in the second gas permeable container. 
     
     
         11 . The method of  claim 1 , wherein expanding the number of TIL comprises culturing the TIL submerged under a height of at least about 2.0 cm of cell culture medium in the second gas permeable container. 
     
     
         12 . The method of  claim 1 , wherein expanding the number of TIL comprises culturing the TIL in a gas permeable container having at least about 650 cm 2  of cell culture surface area. 
     
     
         13 . The method of  claim 1 , wherein expanding the number of TIL comprises increasing the number of TIL by at least about 1000-fold. 
     
     
         14 . The method of  claim 1 , comprising expanding the number of TIL in about 5,000 mL to about 10,000 mL of cell medium. 
     
     
         15 . The method of  claim 1 , wherein expanding the number of TIL uses about 1×10 9  to about 4×10 9  allogeneic feeder cells and/or irradiated autologous feeder cells. 
     
     
         16 . The method of  claim 1 , wherein the cell medium in the first and/or second gas permeable container is unfiltered. 
     
     
         17 . The method of  claim 1 , wherein the cell medium in the first and/or second gas permeable container lacks beta-mercaptoethanol (BME). 
     
     
         18 . The method of  claim 1 , wherein expanding the number of TIL comprises feeding the cells every no more frequently than every third or fourth day. 
     
     
         19 . The method of  claim 1 , wherein the tumor tissue sample is a melanoma tumor tissue sample. 
     
     
         20 . The method of  claim 1 , wherein expanding the number of TIL uses no more than one type of cell culture medium. 
     
     
         21 . The method of  claim 1 , wherein the mammal is a human. 
     
     
         22 . The method of  claim 1 , wherein expanding the number of TIL uses irradiated allogeneic feeder cells and/or irradiated autologous feeder cells at a ratio of about 1 TIL to at least about 20 feeder cells. 
     
     
         23 . The method of  claim 1 , wherein expanding the number of TIL uses irradiated allogeneic feeder cells and/or irradiated autologous feeder cells at a ratio of about 1 TIL to at least about 50 feeder cells. 
     
     
         24 . The method of  claim 1 , wherein expanding the number of TIL uses irradiated allogeneic feeder cells and/or irradiated autologous feeder cells at a ratio of about 1 TIL to at least about 100 feeder cells.