An efficient protocol for the generation of monocyte derived dendritic cells using serum-free media for clinical applications in post remission AML patients

Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on "good manufacture practices" (GMP), have recently been used and see...

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Published in:Annals of clinical and laboratory science Vol. 44; no. 2; p. 180
Main Authors: da Silva Simoneti, Gisele, Saad, Sara Teresinha Olalla, Gilli, Simone Cristina Olenscki
Format: Journal Article
Language:English
Published: United States 2014
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Abstract Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on "good manufacture practices" (GMP), have recently been used and seem to be a viable option. The aim of this study was to evaluate the results of the differentiation, maturation, and function of DCs from Acute Myeloid Leukemia patients (AML), generated in SFM and medium supplemented with autologous serum (AS). DCs were analyzed by phenotype characteristics, viability, and functionality. The results showed the possibility of generating viable DCs in all the conditions tested. In patients, the X-VIVO 15 medium was more efficient than the other media tested in the generation of DCs producing IL-12p70 (p=0.05). Moreover, the presence of AS led to a significant increase of IL-10 by DCs as compared with CellGro (p=0.05) and X-Vivo15 (p=0.05) media, both in patients and donors. We concluded that SFM was efficient in the production of DCs for immunotherapy in AML patients. However, the use of AS appears to interfere with the functional capacity of the generated DCs.
AbstractList Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on "good manufacture practices" (GMP), have recently been used and seem to be a viable option. The aim of this study was to evaluate the results of the differentiation, maturation, and function of DCs from Acute Myeloid Leukemia patients (AML), generated in SFM and medium supplemented with autologous serum (AS). DCs were analyzed by phenotype characteristics, viability, and functionality. The results showed the possibility of generating viable DCs in all the conditions tested. In patients, the X-VIVO 15 medium was more efficient than the other media tested in the generation of DCs producing IL-12p70 (p=0.05). Moreover, the presence of AS led to a significant increase of IL-10 by DCs as compared with CellGro (p=0.05) and X-Vivo15 (p=0.05) media, both in patients and donors. We concluded that SFM was efficient in the production of DCs for immunotherapy in AML patients. However, the use of AS appears to interfere with the functional capacity of the generated DCs.
Author da Silva Simoneti, Gisele
Gilli, Simone Cristina Olenscki
Saad, Sara Teresinha Olalla
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  givenname: Gisele
  surname: da Silva Simoneti
  fullname: da Silva Simoneti, Gisele
  email: mona@unicamp.br
  organization: Instituto Nacional de Ciência e Tecnologia do Sangue, Campinas, São Paulo, 13083-970,Brazil; phone: +55 19 3521 8734; fax: +55 19 3521 8600; e-mail: mona@unicamp.br
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  givenname: Sara Teresinha Olalla
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  fullname: Saad, Sara Teresinha Olalla
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  givenname: Simone Cristina Olenscki
  surname: Gilli
  fullname: Gilli, Simone Cristina Olenscki
BackLink https://www.ncbi.nlm.nih.gov/pubmed/24795057$$D View this record in MEDLINE/PubMed
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Snippet Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease...
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SubjectTerms Adult
Cell Culture Techniques - methods
Cell Differentiation - drug effects
Cell Proliferation - drug effects
Cell Survival - drug effects
Culture Media, Serum-Free - pharmacology
Dendritic Cells - cytology
Dendritic Cells - transplantation
Dextrans - metabolism
Enzyme-Linked Immunosorbent Assay
Female
Flow Cytometry
Fluorescein-5-isothiocyanate - analogs & derivatives
Fluorescein-5-isothiocyanate - metabolism
Humans
Inflammation Mediators - metabolism
Interleukin-10 - biosynthesis
Interleukin-12 - biosynthesis
Leukemia, Myeloid, Acute - therapy
Male
Monocytes - cytology
Phagocytes - cytology
Phagocytes - drug effects
Phenotype
Remission Induction
Tissue Donors
Title An efficient protocol for the generation of monocyte derived dendritic cells using serum-free media for clinical applications in post remission AML patients
URI https://www.ncbi.nlm.nih.gov/pubmed/24795057
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