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Abstract:

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.

Registro:

Documento: Artículo
Título:Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine
Autor:Tapia, I.J.; Aris, M.; Arriaga, J.M.; Blanco, P.A.; Mazzobre, F.; Vega, J.; Mordoh, J.; Barrio, M.M.
Filiación:Centro de Investigaciones Oncológicas FUCA, Ciudad Autónoma de Buenos Aires, Crámer 1180, Primer Piso, CP1426, Argentina
Laboratorio de Propiedades Fisicoquímicas y Conservación de Biomoléculas, Departamento de Industrias, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina
Laboratorio Pablo Cassará, Ciudad Autónoma de Buenos Aires, Caaguazú 7171, CP1408, Argentina
Instituto de Investigaciones Bioquímicas de Buenos Aires, CONICET and Fundación Instituto Leloir, Patricias Argentinas 435 (CP1405), Ciudad Autónoma de Buenos Aires, Argentina
Palabras clave: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
Año:2013
Volumen:67
Número:2
Página de inicio:163
Página de fin:169
DOI: http://dx.doi.org/10.1016/j.cryobiol.2013.06.007
Título revista:Cryobiology
Título revista abreviado:Cryobiology
ISSN:00112240
CODEN:CRYBA
CAS:dimethyl sulfoxide, 67-68-5; human serum albumin, 9048-49-1; trehalose, 99-20-7
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00112240_v67_n2_p163_Tapia

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Citas:

---------- APA ----------
Tapia, I.J., Aris, M., Arriaga, J.M., Blanco, P.A., Mazzobre, F., Vega, J., Mordoh, J.,..., Barrio, M.M. (2013) . Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine. Cryobiology, 67(2), 163-169.
http://dx.doi.org/10.1016/j.cryobiol.2013.06.007
---------- CHICAGO ----------
Tapia, I.J., Aris, M., Arriaga, J.M., Blanco, P.A., Mazzobre, F., Vega, J., et al. "Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine" . Cryobiology 67, no. 2 (2013) : 163-169.
http://dx.doi.org/10.1016/j.cryobiol.2013.06.007
---------- MLA ----------
Tapia, I.J., Aris, M., Arriaga, J.M., Blanco, P.A., Mazzobre, F., Vega, J., et al. "Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine" . Cryobiology, vol. 67, no. 2, 2013, pp. 163-169.
http://dx.doi.org/10.1016/j.cryobiol.2013.06.007
---------- VANCOUVER ----------
Tapia, I.J., Aris, M., Arriaga, J.M., Blanco, P.A., Mazzobre, F., Vega, J., et al. Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine. Cryobiology. 2013;67(2):163-169.
http://dx.doi.org/10.1016/j.cryobiol.2013.06.007