Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine
CSF470 vaccine is a mixture of four lethally irradiated melanoma cell lines, administered with BCG and GM-CSF, which is currently being tested in a Phase II/III Clinical trial in stage II/III melanoma patients. To prepare vaccine doses, irradiated melanoma cell lines are frozen using dimethyl sulfox...
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00112240_v67_n2_p163_Tapia http://hdl.handle.net/20.500.12110/paper_00112240_v67_n2_p163_Tapia |
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paper:paper_00112240_v67_n2_p163_Tapia2023-06-08T14:34:50Z Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine Barrio, María Marcela Cryopreservation Melanoma vaccine Trehalose csf 470 dimethyl sulfoxide human serum albumin liquid nitrogen melanoma vaccine trehalose unclassified drug antigen expression apoptosis article cell death cell level cell proliferation controlled study cryopreservation endocytosis freezing gamma irradiation human human cell low temperature melanoma cell priority journal process optimization thawing Cryopreservation Melanoma vaccine Trehalose Cancer Vaccines Cell Line, Tumor Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Freezing Humans Melanoma Trehalose CSF470 vaccine is a mixture of four lethally irradiated melanoma cell lines, administered with BCG and GM-CSF, which is currently being tested in a Phase II/III Clinical trial in stage II/III melanoma patients. To prepare vaccine doses, irradiated melanoma cell lines are frozen using dimethyl sulfoxide (Me2SO) and stored in liquid nitrogen (liqN2). Prior to inoculation, doses must be thawed, washed to remove Me2SO and suspended for clinical administration. Avoiding the use of Me2SO and storage in liqN2 would allow future freeze-drying of CSF470 vaccine to facilitate pharmaceutical production and distribution. We worked on the development of an alternative cryopreservation methodology while keeping the vaccine's biological and immunogenic properties. We tested different freezing media containing trehalose suitable to remain as excipients in a freeze-dried product, to cryopreserve melanoma cells either before or after gamma irradiation. Melanoma cells incorporated trehalose after 5h incubation at 37°C by fluid-phase endocytosis, reaching an intracellular concentration that varied between 70-140mM depending on the cell line. Optimal freezing conditions were 0.2M trehalose and 30mg/ml human serum albumin, at -84°C. Vaccine doses could be frozen in trehalose at -84°C for at least four months keeping their cellular integrity, antigen expression and apoptosis/necrosis profile after gamma-irradiation as compared to Me2SO control. Non-irradiated melanoma cell lines also showed comparable proliferative capacity after both cryopreservation procedures. Trehalose-freezing medium allowed us to cryopreserve melanoma cells, either alive or after gamma irradiation, at -84°C avoiding the use of Me2SO and liqN2 storage. These cryopreservation conditions could be suitable for future freeze-drying of CSF470 vaccine. © 2013 Elsevier Inc. Fil:Barrio, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2013 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00112240_v67_n2_p163_Tapia http://hdl.handle.net/20.500.12110/paper_00112240_v67_n2_p163_Tapia |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Cryopreservation Melanoma vaccine Trehalose csf 470 dimethyl sulfoxide human serum albumin liquid nitrogen melanoma vaccine trehalose unclassified drug antigen expression apoptosis article cell death cell level cell proliferation controlled study cryopreservation endocytosis freezing gamma irradiation human human cell low temperature melanoma cell priority journal process optimization thawing Cryopreservation Melanoma vaccine Trehalose Cancer Vaccines Cell Line, Tumor Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Freezing Humans Melanoma Trehalose |
| spellingShingle |
Cryopreservation Melanoma vaccine Trehalose csf 470 dimethyl sulfoxide human serum albumin liquid nitrogen melanoma vaccine trehalose unclassified drug antigen expression apoptosis article cell death cell level cell proliferation controlled study cryopreservation endocytosis freezing gamma irradiation human human cell low temperature melanoma cell priority journal process optimization thawing Cryopreservation Melanoma vaccine Trehalose Cancer Vaccines Cell Line, Tumor Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Freezing Humans Melanoma Trehalose Barrio, María Marcela Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| topic_facet |
Cryopreservation Melanoma vaccine Trehalose csf 470 dimethyl sulfoxide human serum albumin liquid nitrogen melanoma vaccine trehalose unclassified drug antigen expression apoptosis article cell death cell level cell proliferation controlled study cryopreservation endocytosis freezing gamma irradiation human human cell low temperature melanoma cell priority journal process optimization thawing Cryopreservation Melanoma vaccine Trehalose Cancer Vaccines Cell Line, Tumor Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Freezing Humans Melanoma Trehalose |
| description |
CSF470 vaccine is a mixture of four lethally irradiated melanoma cell lines, administered with BCG and GM-CSF, which is currently being tested in a Phase II/III Clinical trial in stage II/III melanoma patients. To prepare vaccine doses, irradiated melanoma cell lines are frozen using dimethyl sulfoxide (Me2SO) and stored in liquid nitrogen (liqN2). Prior to inoculation, doses must be thawed, washed to remove Me2SO and suspended for clinical administration. Avoiding the use of Me2SO and storage in liqN2 would allow future freeze-drying of CSF470 vaccine to facilitate pharmaceutical production and distribution. We worked on the development of an alternative cryopreservation methodology while keeping the vaccine's biological and immunogenic properties. We tested different freezing media containing trehalose suitable to remain as excipients in a freeze-dried product, to cryopreserve melanoma cells either before or after gamma irradiation. Melanoma cells incorporated trehalose after 5h incubation at 37°C by fluid-phase endocytosis, reaching an intracellular concentration that varied between 70-140mM depending on the cell line. Optimal freezing conditions were 0.2M trehalose and 30mg/ml human serum albumin, at -84°C. Vaccine doses could be frozen in trehalose at -84°C for at least four months keeping their cellular integrity, antigen expression and apoptosis/necrosis profile after gamma-irradiation as compared to Me2SO control. Non-irradiated melanoma cell lines also showed comparable proliferative capacity after both cryopreservation procedures. Trehalose-freezing medium allowed us to cryopreserve melanoma cells, either alive or after gamma irradiation, at -84°C avoiding the use of Me2SO and liqN2 storage. These cryopreservation conditions could be suitable for future freeze-drying of CSF470 vaccine. © 2013 Elsevier Inc. |
| author |
Barrio, María Marcela |
| author_facet |
Barrio, María Marcela |
| author_sort |
Barrio, María Marcela |
| title |
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| title_short |
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| title_full |
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| title_fullStr |
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| title_full_unstemmed |
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine |
| title_sort |
development of a novel methodology for cryopreservation of melanoma cells applied to csf470 therapeutic vaccine |
| publishDate |
2013 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00112240_v67_n2_p163_Tapia http://hdl.handle.net/20.500.12110/paper_00112240_v67_n2_p163_Tapia |
| work_keys_str_mv |
AT barriomariamarcela developmentofanovelmethodologyforcryopreservationofmelanomacellsappliedtocsf470therapeuticvaccine |
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1768543638632005632 |