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Processing of tumor tissues for vaccination with autologous tumor cells
1Abteilungen Innere Medizin I, Hämatologie/Onkologie, Universitätsklinik Freiburg, Deutschland.
Abstract:
Vaccination with gene-transfected tumor cells has recently been proposed as a new strategy in the immunotherapy of cancer. Since autologous tumor cells provide an optimal antigen profile, the possibility of generating single cell suspensions from renal cell carcinoma (RCC), malignant melanoma (MM), colon carcinoma (CC), and non-small-cell lung cancer (NSCLC) biopsies was investigated. One hundred and seventy-four tumor biopsies were processed by mechanic and enzymatic dissociation, yielding 1-2 x 10(6) cells/g tumor (median), irrespective of tumor type. Primary tumor cell cultures (PTCC) of > or = 10(7) cells were established from 29 of 86 (34%) RCC, 14 of 38 (37%) MM, 11 of 23 (48%) NSCLC and 4 of 27 (15%) CC specimens. The amount of non-tumor cells, as assessed by morphology and immunocytology, was generally low (< 30%) in RCC (35 of 41) and MM (11 of 17), while it exceeded 60% in 8 of 11 PTCC from NSCLC and 3 of 11 CC. A high tumor cell yield was obtained in biopsies with a high degree of vascularization and in the virtual absence of necrosis. Thus, PTCC > or = 10(7) cells were obtained in 73% of MM with a high degree of vascularization and in 22% of MM with a low degree of vascularization (p < 0.007). Long-term tumor cell cultures exceeding 20 passages were established in 24 of 86 (18%) RCC, 7 of 38 (18%) MM and 3 of 27 (11%) CC, while successful implantation in nude mice was achieved in 8 of 20 RCC and 5 of 10 MM. Thus, under the conditions described, > or = 10(7) primary tumor cells of high purity could be generated from about one third of RCC and MM biopsies, while the success rate increased to > 50 and > 70%, respectively, in samples with a high degree of vascularization generated by an optimized biopsy technique excluding necrotic parts.
Insights
Generating sufficient pure primary tumor cell cultures (PTCC) for cancer immunotherapy is feasible. Optimized biopsy techniques improve cell yield and purity, especially for renal cell carcinoma and malignant melanoma, enhancing potential for gene-transfected tumor cell vaccines.
Area of Science:
- Oncology
- Immunotherapy
- Cell Biology
Background:
- Gene-transfected tumor cell vaccination is a novel cancer immunotherapy strategy.
- Autologous tumor cells offer an optimal antigen profile for personalized treatment.
- Generating sufficient viable tumor cells from patient biopsies is crucial for this approach.
Purpose of the Study:
- To investigate the feasibility of generating single-cell suspensions for primary tumor cell cultures (PTCC).
- To assess the yield and purity of PTCC from various cancer types: renal cell carcinoma (RCC), malignant melanoma (MM), colon carcinoma (CC), and non-small-cell lung cancer (NSCLC).
- To identify factors influencing successful PTCC generation for immunotherapy applications.
Main Methods:
- Mechanic and enzymatic dissociation of 174 tumor biopsies.
- Establishment and culture of PTCC, aiming for > or = 10(7) cells.
- Assessment of non-tumor cell content via morphology and immunocytology.
- Evaluation of tumor vascularization and necrosis impact on cell yield.
- Long-term culture viability and nude mouse implantation studies.
Main Results:
- PTCC of > or = 10(7) cells were established from 34% RCC, 37% MM, 48% NSCLC, and 15% CC.
- Low non-tumor cell content (<30%) was typical for RCC and MM.
- High tumor cell yield correlated significantly with high biopsy vascularization and minimal necrosis (e.g., 73% success in highly vascularized MM).
- Long-term cultures (>20 passages) and successful nude mouse implantation were achieved in subsets of RCC and MM.
Conclusions:
- Generating > or = 10(7) high-purity primary tumor cells is achievable from approximately one-third of RCC and MM biopsies.
- Optimized biopsy techniques, focusing on vascularity and excluding necrotic tissue, significantly increase success rates.
- These findings support the potential of using autologous, gene-transfected tumor cells for cancer immunotherapy.