کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10928054 | 1092891 | 2013 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Development of a novel methodology for cryopreservation of melanoma cells applied to CSF470 therapeutic vaccine
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم کشاورزی و بیولوژیک
علوم کشاورزی و بیولوژیک (عمومی)
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چکیده انگلیسی
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 5 h incubation at 37 °C by fluid-phase endocytosis, reaching an intracellular concentration that varied between 70-140 mM depending on the cell line. Optimal freezing conditions were 0.2 M trehalose and 30 mg/ml human serum albumin, at â84 °C. Vaccine doses could be frozen in trehalose at â84 °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 â84 °C avoiding the use of Me2SO and liqN2 storage. These cryopreservation conditions could be suitable for future freeze-drying of CSF470 vaccine.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Cryobiology - Volume 67, Issue 2, October 2013, Pages 163-169
Journal: Cryobiology - Volume 67, Issue 2, October 2013, Pages 163-169
نویسندگان
Ivana J. Tapia, Mariana Aris, Juan MartÃn Arriaga, Paula A. Blanco, Florencia Mazzobre, Julio Vega, José Mordoh, MarÃa Marcela Barrio,