Retrospective Assessment of Clonality of a Legacy Cell Line by Analytical Subcloning of the Master Cell Bank

Abstract Background: Cell banks have been widely used to preserve cell properties as well as to record and control cell line access in research. However, the generation of cell banks involved in the manufacturing of Advanced therapy medicinal products such as cell or gene therapy products must comply with the current Good Manufacturing Practice regulation. Similarly, the quality of those cell lines used as starting materials in viral-vector manufacturing processes must be also evaluated.Methods: Three batches of both Master Cell Bank and Working Cell Bank of the HEK293T cell line were manufactured under the current Good Manufacturing Practices regulation. Quality control test were performed according to the product specifications. The process validation includes previous qualification of the manufacturing personnel by performing simulation tests as well as the continuous measure of environmental parameters during manufacturing such as air particles and microorganism. Cell number and viability of cryopreserved cells were periodically measured in order to define the stability of these cellular products.Results: All batches of Master Cell Bank and Working Cell Bank fulfilled the acceptance criteria of their specifications showing the robustness and homogeneity of the processes. In addition, both Master and Working cell bank maintain the defined viability and cell number 37 months after cryopreservation. Conclusions: Manufacturing cell banks under Good Manufacturing Practices regulation for its use as raw material or final cellular product is feasible. HEK293T cell banks have been used to manufacture clinical-grade lentiviral particles for Chimeric Antigen Receptor T-cell based clinical trials.

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Cell Banking of HEK293T Cell Line for Clinical-Grade Lentiviral Particles Manufacturing

10.21203/rs.3.rs-54098/v1 ◽

2020 ◽

Author(s):

Unai Perpiña ◽

Cristina Herranz ◽

Raquel Martin-Ibañez ◽

Anna Boronat ◽

Felipe Chiappe ◽

...

Keyword(s):

Cell Line ◽

Hek293t Cell ◽

Cell Number ◽

Clinical Grade ◽

Cell Bank ◽

Master Cell Bank ◽

Good Manufacturing Practices ◽

Working Cell ◽

Cell Banks ◽

Hek293t Cell Line

Abstract Background: Cell banks have been widely used to preserve cell properties as well as to record and control cell line access in research. However, the generation of cell banks involved in the manufacturing of Advanced therapy medicinal products such as cell or gene therapy products must comply with the current Good Manufacturing Practice regulation. Similarly, the quality of those cell lines used as starting materials in viral-vector manufacturing processes must be also evaluated.Methods: Three batches of both Master Cell Bank and Working Cell Bank of the HEK293T cell line were manufactured under the current Good Manufacturing Practices regulation. Quality control test were performed according to the product specifications. The process validation includes previous qualification of the manufacturing personnel by performing simulation tests as well as the continuous measure of environmental parameters during manufacturing such as air particles and microorganism. Cell number and viability of cryopreserved cells were periodically measured in order to define the stability of these cellular products.Results: All batches of Master Cell Bank and Working Cell Bank fulfilled the acceptance criteria of their specifications showing the robustness and homogeneity of the processes. In addition, both Master and Working cell bank maintain the defined viability and cell number 37 months after cryopreservation. Conclusions: Manufacturing cell banks under Good Manufacturing Practices regulation for its use as raw material or final cellular product is feasible. HEK293T cell banks have been used to manufacture clinical-grade lentiviral particles for Chimeric Antigen Receptor T-cell based clinical trials.

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Cell Banking of HEK293T cell line for clinical-grade lentiviral particles manufacturing

10.21203/rs.3.rs-71411/v2 ◽

2020 ◽

Author(s):

Unai Perpiña ◽

Cristina Herranz ◽

Raquel Martin-Ibañez ◽

Anna Boronat ◽

Felipe Chiappe ◽

...

Keyword(s):

Cell Line ◽

Cell Lines ◽

Hek293t Cell ◽

Good Manufacturing Practice ◽

Clinical Grade ◽

Cell Bank ◽

Advanced Therapy ◽

Working Cell ◽

Cell Banks ◽

Hek293t Cell Line

Abstract Background: Cell banks are widely used to preserve cell properties as well as to record and control the use of cell lines in biomedical research. The generation of cell banks for the manufacturing of Advanced Therapy Medicinal Products, such as cell and gene therapy products, must comply with current Good Manufacturing Practice regulations. The quality of the cell lines used as starting materials in viral-vector manufacturing processes must be also assessed.Methods: Three batches of a Master Cell Bank and a Working Cell Bank of the HEK293T cell line were manufactured under current Good Manufacturing Practices regulations. Quality control tests were performed according to product specifications. Process validation includes the training of manufacturing personnel by performing simulation tests, and the continuous measurement of environmental parameters such as air particles and microorganisms. Cell number and viability of cryopreserved cells were periodically measured in order to define the stability of these cellular products.Results: All batches of HEK293T Master and Working Cell Banks met the acceptance criteria of their specifications showing the robustness and homogeneity of the processes. In addition, both Master and Working Cell Banks maintained the defined cell viability and concentration over a 37 month-period after cryopreservation. Conclusions: Manufacturing cell banks under Good Manufacturing Practice regulations for their use as raw materials or final cellular products is feasible. HEK293T cell banks were used to manufacture clinical-grade lentiviral particles for Chimeric Antigen Receptor T-cell based clinical trials.

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Genetic variability in a frozen batch of MCF-7 cells invisible in routine authentication affecting cell function

Scientific Reports ◽

10.1038/srep28994 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 42

Author(s):

Andre Kleensang ◽

Marguerite M. Vantangoli ◽

Shelly Odwin-DaCosta ◽

Melvin E. Andersen ◽

Kim Boekelheide ◽

...

Keyword(s):

Cell Line ◽

Genetic Heterogeneity ◽

Cell Function ◽

Breast Adenocarcinoma ◽

Comparative Genomic ◽

Cell Bank ◽

Cell Line Authentication ◽

Str Markers ◽

Genetic Drifts ◽

Mcf 7

Abstract Common recommendations for cell line authentication, annotation and quality control fall short addressing genetic heterogeneity. Within the Human Toxome Project, we demonstrate that there can be marked cellular and phenotypic heterogeneity in a single batch of the human breast adenocarcinoma cell line MCF-7 obtained directly from a cell bank that are invisible with the usual cell authentication by short tandem repeat (STR) markers. STR profiling just fulfills the purpose of authentication testing, which is to detect significant cross-contamination and cell line misidentification. Heterogeneity needs to be examined using additional methods. This heterogeneity can have serious consequences for reproducibility of experiments as shown by morphology, estrogenic growth dose-response, whole genome gene expression and untargeted mass-spectroscopy metabolomics for MCF-7 cells. Using Comparative Genomic Hybridization (CGH), differences were traced back to genetic heterogeneity already in the cells from the original frozen vials from the same ATCC lot, however, STR markers did not differ from ATCC reference for any sample. These findings underscore the need for additional quality assurance in Good Cell Culture Practice and cell characterization, especially using other methods such as CGH to reveal possible genomic heterogeneity and genetic drifts within cell lines.

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Screening for 15 pathogenic viruses in human cell lines registered at the JCRB Cell Bank: characterization of in vitro human cells by viral infection

Royal Society Open Science ◽

10.1098/rsos.172472 ◽

2018 ◽

Vol 5 (5) ◽

pp. 172472 ◽

Cited By ~ 2

Author(s):

Setsuko Shioda ◽

Fumio Kasai ◽

Ken Watanabe ◽

Kohei Kawakami ◽

Azusa Ohtani ◽

...

Keyword(s):

Viral Infection ◽

Cell Line ◽

Cell Lines ◽

Human Cell ◽

The Other ◽

Pcr Analysis ◽

Human Cell Lines ◽

Cell Bank

Human cell lines have been used in a variety of research fields as an in vitro model. These cells are all derived from human tissue samples, thus there is a possibility of virus infection. Virus tests are routinely performed in clinical practice, but are limited in cell lines. In this study, we investigated 15 kinds of viruses in 844 human cell lines registered at the Japanese Collection of Research Bioresources (JCRB) Cell Bank. Our real-time PCR analysis revealed that six viruses, EBV, HTLV-1, HBV, B19V, HHV-6 and HHV-7, were detected in 43 cell lines. Of them, 20 cell lines were transformed by intentional infection in vitro with EBV or HTLV-1. Viruses in the other 23 cell lines and one EBV transformed cell line are derived from an in vivo infection, including five de novo identifications of EBV, B19V or HHV-7 carriers. Among them, 17 cell lines were established from patients diagnosed with virus-associated diseases. However, the other seven cell lines originated from in vivo cells unrelated to disease or cellular tropism. Our approach to screen for a set of 15 viruses in each cell line has worked efficiently to identify these rare cases. Virus tests in cell lines contribute not only to safety assessments but also to investigation of in vivo viral infection which can be a characteristic feature of cell lines.

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Cost-Effective Master Cell Bank Validation of Multiple Clinical-Grade Human Pluripotent Stem Cell Lines From a Single Donor

Stem Cells Translational Medicine ◽

10.5966/sctm.2014-0015 ◽

2014 ◽

Vol 3 (10) ◽

pp. 1116-1124 ◽

Cited By ~ 12

Author(s):

Liani Devito ◽

Anastasia Petrova ◽

Cristian Miere ◽

Stefano Codognotto ◽

Nicola Blakely ◽

...

Keyword(s):

Stem Cell ◽

Cell Lines ◽

Pluripotent Stem Cell ◽

Cost Effective ◽

Clinical Grade ◽

Human Pluripotent Stem Cell ◽

Cell Bank ◽

Single Donor ◽

Master Cell Bank ◽

Stem Cell Lines

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Root cause investigation of a viral contamination incident occurred during master cell bank (MCB) testing and characterization – A case study

Biologicals ◽

10.1016/j.biologicals.2008.07.005 ◽

2008 ◽

Vol 36 (6) ◽

pp. 393-402 ◽

Cited By ~ 11

Author(s):

Dayue Chen ◽

Raymond Nims ◽

Sandra Dusing ◽

Pamela Miller ◽

Wen Luo ◽

...

Keyword(s):

Cell Bank ◽

Viral Contamination ◽

Root Cause ◽

Master Cell Bank

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Establishing a master cell bank of bone marrow-derived mesenchymal stem cells for the cellular immunotherapy for septic shock trial (CISS)

Cytotherapy ◽

10.1016/j.jcyt.2015.03.478 ◽

2015 ◽

Vol 17 (6) ◽

pp. S50

Author(s):

Shirley H. Mei ◽

Mahmoud Salkhordeh ◽

Saad Khan ◽

Samantha Hodgins ◽

Irene Watpool ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Septic Shock ◽

Mesenchymal Stem Cells ◽

Cellular Immunotherapy ◽

Shock Trial ◽

Cell Bank ◽

Master Cell Bank

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Improving Biologics’ Effectiveness in Clinical Oncology: From the Combination of Two Monoclonal Antibodies to Oligoclonal Antibody Mixtures

Cancers ◽

10.3390/cancers13184620 ◽

2021 ◽

Vol 13 (18) ◽

pp. 4620

Author(s):

Christel Larbouret ◽

Laurent Gros ◽

André Pèlegrin ◽

Thierry Chardès

Keyword(s):

Clinical Trials ◽

Monoclonal Antibodies ◽

Resistance Mechanisms ◽

Therapeutic Effects ◽

Cell Bank ◽

Humoral Immune ◽

Cancer Management ◽

Biological Responses ◽

Master Cell Bank ◽

Single Batch

Monoclonal antibodies have revolutionized the treatment of many diseases, but their clinical efficacy remains limited in some other cases. Pre-clinical and clinical trials have shown that combinations of antibodies that bind to the same target (homo-combinations) or to different targets (hetero-combinations) to mimic the polyclonal humoral immune response improve their therapeutic effects in cancer. The approval of the trastuzumab/pertuzumab combination for breast cancer and then of the ipilimumab/nivolumab combination for melanoma opened the way to novel antibody combinations or oligoclonal antibody mixtures as more effective biologics for cancer management. We found more than 300 phase II/III clinical trials on antibody combinations, with/without chemotherapy, radiotherapy, small molecules or vaccines, in the ClinicalTrials.gov database. Such combinations enhance the biological responses and bypass the resistance mechanisms observed with antibody monotherapy. Usually, such antibody combinations are administered sequentially as separate formulations. Combined formulations have also been developed in which separately produced antibodies are mixed before administration or are produced simultaneously in a single cell line or a single batch of different cell lines as a polyclonal master cell bank. The regulation, toxicity and injection sequence of these oligoclonal antibody mixtures still need to be addressed in order to optimize their delivery and their therapeutic effects.

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Integrated Collection of Stem Cell Bank data, a data portal for standardized stem cell information

10.1101/2020.08.26.263830 ◽

2020 ◽

Author(s):

Ying Chen ◽

Kunie Sakurai ◽

Sumihiro Maeda ◽

Tohru Masui ◽

Hideyuki Okano ◽

...

Keyword(s):

Stem Cell ◽

Cell Line ◽

Cell Lines ◽

Data Exchange ◽

Database Search ◽

Stem Cell Line ◽

Cell Bank ◽

Data Portal ◽

Stem Cell Lines ◽

Major Data

SUMMARYThe last decade has witnessed an extremely rapid increase in the number of newly established stem cell lines worldwide. However, due to the lack of a standardized format, data exchange among stem cell line information resources has been challenging, and no system can search all stem cell lines across resources worldwide. To solve this problem, we have developed the Integrated Collection of Stem Cell Bank data (ICSCB) (http://icscb.stemcellinformatics.org/), a new and largest database search portal for stem cell line data from various resources, based on the standardized data items and terms of the MIACARM framework. Currently, ICSCB can retrieve >16,000 cell lines from four major data resources in Europe, Japan, and the U.S. ICSCB is automatically updated to provide the latest cell line information, and its integrative search engine helps users collect cell line information from donors with over 1,000 diseases including many rare diseases worldwide, which has been a formidable task, thereby distinguishing itself from other database search portals.

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