Abstract: The present disclosure provides methods for genetically engineering T cells, such as CD4+ T cells, for use in cell therapy. In some aspects, the provided methods include one or more steps for incubating the cells under stimulating conditions, introducing a recombinant polypeptide to the cells through transduction or transfection, and cultivating the cells under conditions that promote proliferation and/or expansion. In some aspects, the incubation and/or the cultivation is performed in the presence of recombinant IL-2. In some aspects, the provided methods are an efficient, reliable means to produce genetically engineered T cells with a high degree of success.

Abstract: The present disclosure is directed to closed-system cryogenic vessels for biomedical material with needleless removal. The needleless removal can reduce damage to biomedical material inside the vessel, allow for greater recovery of biomedical material from the vessel, and reduce exposure risk to users of the closed-system cryogenic vessels during removal of the biomedical material from the vessel. In some aspects, the vessels can be used to store or package a composition of cells, such as a composition containing engineered cells, including in connection with adoptive cell therapy. Also provided are articles of manufacture, kits and methods.