We could therefore exert that PB is stable up to 48 h from harvesting and this allows to wait for the virologic test results before starting the expansion process. From the comparative study, we observed no differences in terms of viability, cellular growth and identity between CIKs that expanded from fresh PB and those processed at 24 and 48 h from collection (data not shown). We first tested PB stability at 24 and 48 h from donor’s harvesting. Įach expansion run started from peripheral blood (PB) and ended with the evaluation of cellular viability, growth in terms of fold increase, identity and cytotoxicity.Īs GMP guidelines recommend, blood has to be kept in quarantine in a blood bank refrigerator until the results of virology and sterility tests are provided. FBS was used as a control condition as our preclinical data were obtained culturing CIKs with RPMI + 10% FBS. In order to replace FBS, commonly used for CIKs expansion, we decided to test Human Serum (HS) and Human Pool Plasma (HPP) as supplement medium. The aim of the study was to validate a GMP method for CIKs production as ATMP for treatment of human sarcomas. Īim, study design and setting of the study Moreover, according to GMP guidelines imposing to provide acceptance criteria for each pharmaceutical ATMP, and on the bases of our preclinical data, we have established that CIKs were compliant when viability was > 80%, CD3+ > 80% ± 10%, CD3+CD56+ ≥ 15%. GMP ensures that ATMP are consistently produced and controlled to the quality standards required to their intended use, from the collection and manipulation of raw materials to the processing of intermediate products, quality controls, storage, labelling and packaging, and release.
As reported in Directive 1394/2007, in which “the principles and guidelines of GMP in respect of medicinal products for human use and investigational medicinal products for human use” are defined, the safest and the most advantageous CIKs expansion method that will be validated. On these bases, our aim was translating the expansion method from preclinical to GMP field. This is due to the xenogenic sera high batch-to-batch variability and to the associated risk of transmitting infectious agents. Therefore, regulatory guidelines for Good Manufacturing Practices (GMP) production, aimed to minimize FBS, used in most ex vivo expansion protocols as medium supplement. CIK cells with CD3+CD56+ immunophenotype have cytotoxic effects similar to those previously obtained by other groups, as they efficiently killed sarcoma cells. performed preliminary study in which CIKs were expanded in RPMI + 10% Fetal Bovine Serum (FBS) to evaluate their effectiveness for antitumor activity against autologous bone sarcomas. They can be classically expanded starting from peripheral blood mononuclear cells (PBMCs) but may also be generated from bone marrow (BM) or umbilical cord blood precursors. The clinical translation of adoptive immunotherapy with CIK cells as treatment for patients with solid tumors is currently the object of clinical trials and their number has increased in recent years. In conclusion, these preclinical validation data lay the bases for a GMP-compliant process to improve the CIKs expansion method.Ĭytokine-induced killer (CIK) cells are a very promising cell population which raise growing interest in the field of cellular antitumor therapy mainly due to their easy expansion method. We performed a validation consisting in three run-sand even if the small number of experiments didn’t permit us to obtained statistical results we demonstrated that both X-VIVO 15 and Tex Macs fulfilled the quality standards in terms of cellular growth, viability and identity while Cell Genix GMP SCGM resulted not compliant as it caused some technical problems such as high mortality.
#Texmacs gmp medium free#
Consequently, we decided to test three different serum free expansion media: X-VIVO 15, (largely used by other groups) and Tex Macs and Cell Genix GMP SCGM: two GMP manufactured media. Firstly, Human Serum (HS) and Human Pool Plasma (HPP) were tested as medium supplements giving not compliant results to acceptance criteria, established for CIKs, probably for the great batch to batch variability. We decided to replace Fetal Bovine Serum (FBS), largely used as medium supplement for CIKs expansion, with other culture media. For this reason, the use of the xenogenic sera tended to be minimized by GMP for their high variability and the associated risk of transmitting infectious agents. GMP ensures that ATMP are consistently produced and controlled to the quality standards required to their intended use. The aim of our study was to validate the most advantageous expansion method for this advanced therapy medicinal product (ATMP) and to translate it from preclinical field to good manufacturing practices (GMP). Cytokine-induced killer (CIK) cells are a very promising cell population raising growing interest in the field of cellular antitumor therapy.
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