Development of an optimized lentiviral transduction medium and process to manufacture genetically modified MSC working cell banks

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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Safety and Immunogenicity of Single-Dose Live Oral Cholera Vaccine Strain CVD 103-HgR, Prepared from New Master and Working Cell Banks

Clinical and Vaccine Immunology ◽

10.1128/cvi.00601-13 ◽

2013 ◽

Vol 21 (1) ◽

pp. 66-73 ◽

Cited By ~ 32

Author(s):

Wilbur H. Chen ◽

Richard N. Greenberg ◽

Marcela F. Pasetti ◽

Sofie Livio ◽

Michael Lock ◽

...

Keyword(s):

United States ◽

Single Dose ◽

Cholera Toxin ◽

Vaccine Strain ◽

The United States ◽

Cholera Vaccine ◽

Oral Cholera Vaccine ◽

Working Cell ◽

Cell Banks ◽

Placebo Recipients

ABSTRACTCurrently, no cholera vaccine is available for persons traveling from the United States to areas of high cholera transmission and who for reasons of occupation or host factors are at increased risk for development of the disease. A single-dose oral cholera vaccine with a rapid onset of protection would be particularly useful for such travelers and might also be an adjunct control measure for cholera outbreaks. The attenuatedVibrio choleraeO1 vaccine strain CVD 103-HgR harbors a 94% deletion of the cholera toxin A subunit gene (ctxA) and has a mercury resistance gene inserted in the gene encoding hemolysin A. We undertook a phase I randomized placebo-controlled two-site trial to assess the safety and immunogenicity of a preliminary formulation of CVD 103-HgR prepared from new master and working cell banks. Healthy young adults were randomized (5:1 vaccinees to placebo recipients) to receive a single oral dose of ∼4.4 × 108CFU of vaccine or a placebo. Blood serum vibriocidal and cholera toxin-specific IgG antibodies were measured before and 10, 14, and 28 days following vaccination or placebo. Excretion of the vaccine strain in the stool was assessed during the first week postvaccination. A total of 66 subjects were enrolled, comprising 55 vaccinees and 11 placebo recipients. The vaccine was well tolerated. The overall vibriocidal and anti-cholera toxin seroconversion rates were 89% and 57%, respectively. CVD 103-HgR is undergoing renewed manufacture for licensure in the United States under the auspices of PaxVax. Our data mimic those from previous commercial formulations that elicited vibriocidal antibody seroconversion (a correlate of protection) in ∼90% of vaccinees. (This study has been registered at ClinicalTrials.gov under registration no. NCT01585181.)

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

10.21203/rs.3.rs-71411/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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Development of a Novel Thermoplastic Tubing, FP-FLEX™, and Single-Use Freezing Bag for Working Cell Banks Enabling Closed-System Processing to Temperatures as Low as -196°C

BMC Proceedings ◽

10.1186/1753-6561-9-s9-p58 ◽

2015 ◽

Vol 9 (S9) ◽

Author(s):

Dominic Clarke ◽

Doug Norris ◽

Bradley Bennett

Keyword(s):

Closed System ◽

Working Cell ◽

Single Use ◽

Cell Banks

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Lentiviral transduction of neonatal rat ventricular myocytes preserves ultrastructural features of genetically modified cells

Virology ◽

10.1016/j.virol.2021.07.015 ◽

2021 ◽

Author(s):

S.R. Consonni ◽

A.C.P.V. de Carvalho ◽

B.A. Mauro ◽

K.G. Franchini ◽

M.C. Bajgelman

Keyword(s):

Ventricular Myocytes ◽

Genetically Modified ◽

Neonatal Rat ◽

Lentiviral Transduction ◽

Rat Ventricular Myocytes ◽

Neonatal Rat Ventricular Myocytes

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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

Translational Medicine Communications ◽

10.1186/s41231-020-00075-w ◽

2020 ◽

Vol 5 (1) ◽

Author(s):

Unai Perpiñá ◽

Cristina Herranz ◽

Raquel Martín-Ibáñez ◽

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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