Cryopreservation of adenovirus-transfected dendritic cells (DCs) for clinical use

D. Gülen, S. Maas, H. Julius, Phyllis Irene Warkentin, H. Britton, I. Younos, J. Senesac, Samuel Jay Pirruccello, James E Talmadge

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

In this study, we examined the effects of cryoprotectant, freezing and thawing, and adenovirus (Adv) transduction on the viability, transgene expression, phenotype, and function of human dendritic cells (DCs). DCs were differentiated from cultured peripheral blood (PB) monocytes following Elutra isolation using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 6 days and then transduced using an Adv vector with an IL-12 transgene. Fresh, cryopreserved, and thawed transduced immature DCs were examined for their: 1) cellular concentration and viability; 2) antigenicity using an allogeneic mixed lymphocyte reaction (MLR); 3) phenotype (HLA-DR and CD11c) and activation (CD83); and 4) transgene expression based on IL-12 secretion. Stability studies revealed that transduced DCs could be held in cryoprotectant for as long as 75 min at 2-8 °C prior to freezing with little effect on their viability and cellularity. Further, cryopreservation, storage, and thawing reduced the viability of the transduced DCs by an average of 7.7%; and had no significant impact on DC phenotype and activation. In summary, cryopreservation, storage, and thawing had no significant effect on DC viability, function, and transgene expression by Adv-transduced DCs.

Original languageEnglish (US)
Pages (from-to)61-68
Number of pages8
JournalInternational Immunopharmacology
Volume13
Issue number1
DOIs
StatePublished - May 1 2012

Fingerprint

Cryopreservation
Adenoviridae
Dendritic Cells
Transgenes
Interleukin-12
Phenotype
Freezing
Mixed Lymphocyte Culture Test
HLA-DR Antigens
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-4
Monocytes
Interleukin-6
Cell Survival

Keywords

  • Adenovirus
  • Cryopreservation
  • Dendritic cells
  • Stability
  • Viability

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Pharmacology

Cite this

Cryopreservation of adenovirus-transfected dendritic cells (DCs) for clinical use. / Gülen, D.; Maas, S.; Julius, H.; Warkentin, Phyllis Irene; Britton, H.; Younos, I.; Senesac, J.; Pirruccello, Samuel Jay; Talmadge, James E.

In: International Immunopharmacology, Vol. 13, No. 1, 01.05.2012, p. 61-68.

Research output: Contribution to journalArticle

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