Immunotherapy using IL-2 alone or combined with activated lymphocytes has been promising for metastatic renal cell carcinoma. Cytotoxic lymphocytes can be isolated from tumors, expanded in vitro with IL-2, and adoptively transferred back into the tumor-bearing host. These cells can also be transduced with the genes coding for cytokines for local delivery to tumor sites. A major drawback in adoptive immunotherapy is the cumbersome and expensive culture technology associated with the growth of large numbers of cells required for their therapeutic effect. To reduce the cost, resources, and manpower, we have developed the methodology for lymphocyte activation and expansion in the automated hollow fiber bioreactor IMMUNO*STAR™ Cell Expander (ACT BIOMEDICAL, INC). Tumor Infiltrating Lymphocytes (TIL) isolated from human renal cell carcinoma tumor specimens were inoculated at a number of 108 cells in a small bioreactor of 30 ml extracapillary space volume. We have determined the medium flow rates and culture conditions to obtain a significant and repeated expansion of TIL at weekly intervals. The lymphocytes cultured in the bioreactor demonstrated the same phenotype and cytotoxic activity as those expanded in parallel in tissue culture plates. Lymphocyte expansion in the hollow fiber bioreactor required lower volumes of medium, human serum, IL-2 and minimal labor. This technology may facilitate the use of adoptive immunotherapy for the treatment of refractory malignancies.

01 Jan 1994·Animal Cell Technology: Basic & Applied Aspects

Cryopreservation of Cell-Laden Collagen Microspheres

Author: Aline Nadeau ; Amar Tung ; Akihiro Nishi ; Ernest A. Bognar ; Thomas C. Ripley

Cryopreservation of viable cells is one of biotechnology’s fundamental building blocks. From research through development to commercialization of biopharmaceuticals, the ability to bank cells for future use is a critical step to validate a given process. Although guidelines for the creation and testing of both master cell banks (MCB) and manufacturers working cell banks (MWCB) are numerous, a trend of laying down cell banks in conditions as close to the manufacturing process as feasible is being formed. Also, the ability to cryopreserve cells from the production phase of the manufacturing process is becoming an increasingly important tool in validating continuous culture systems, especially for attachment dependent cell types. With this frame of reference, we have demonstrated the cryopreservation of cell-laden collagen microspheres with both attachment dependent and suspension cell types. Discussed are procedures, cell types, acceptance criteria, analyses, and planned future work including applications in large scale cell banking.

01 May 1993·American biotechnology laboratory

A synthetic affinity ligand for purifying immunoglobulins from goat serum.

Article

Author: Ngo, T ; Van Sluis, R ; Rosenberg, J

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