PMID- 23269574
OWN - NLM
STAT- MEDLINE
DCOM- 20130516
LR  - 20211021
IS  - 1096-0945 (Electronic)
IS  - 0014-4800 (Print)
IS  - 0014-4800 (Linking)
VI  - 94
IP  - 2
DP  - 2013 Apr
TI  - Generation of large numbers of highly purified dendritic cells from bone marrow 
      progenitor cells after co-culture with syngeneic murine splenocytes.
PG  - 336-42
LID - S0014-4800(12)00176-1 [pii]
LID - 10.1016/j.yexmp.2012.12.001 [doi]
AB  - Dendritic cells (DCs) are called the sentinels of the human immune system because 
      of their function as antigen presenting cells (APCs) that elicit a protective 
      immune response. Given that DCs have been used for many years as target cells in 
      a great number of experiments, it became essential to devise a new method for 
      producing DCs in higher quantities and of greater purity. Here we report a novel 
      technique for obtaining more dendritic cells, and with higher purity, from 
      in-vitro co-culture of bone marrow (BM) cells with splenocytes. From a total of 
      20 x 10(6) BM cells and 120 x 10(6)splenocytes, 3 x 10(6) BM cells along with 20 
      x 10(6)splenocytes were co-cultured in petri dishes for DC generation; 120 x 
      10(6) splenocytes from one C57BL/6 mouse were also co-cultured in petri dishes 
      for DC generation. BM cells were the control group cultured in the same 
      conditions except for the presence of splenocytes. Purity and maturation state of 
      DCs were checked by lineage surface markers (CD11c, CD11b, CD8alpha, and F4/80) and 
      the expression levels of MHCII as well as co-stimulatory molecules (CD86, CD80, 
      and CD40). Endocytosis and thymidine uptake capacity were also used to test the 
      functionality of DCs. The levels of IL-12p70, IL-23, and IL-10 were also checked 
      in the supernatant of cultured cells by ELISA. The number of DCs derived from 
      co-culture of BM and splenocytes (DCs(TME)) was at least twice that of BM-derived 
      DCs in the absence of splenocytes. In addition, the purity of DCs after 
      co-culture of BM and splenocytes was greater than that of DCs in the control 
      culture (90.2% and 77.2%, respectively; p<0.05). While functional assays showed 
      no differences between co-culture and control groups, IL-10 levels were 
      significantly lower in DCs(TME) compared to BM-derived DCs in the absence of 
      splenocytes (193 pg/ml and 630 pg/ml, respectively; p<0.05). The results of the 
      present study show that the generation of DCs from BM progenitors is accelerated 
      in the presence of syngeneic splenocytes. Given the larger number of generated 
      DCs, and with higher purity, in this technique, DCs(TME) could be more 
      advantageous for DC-based immunotherapy and vaccination techniques.
CI  - Copyright (c) 2012 Elsevier Inc. All rights reserved.
FAU - Kalantari, Tahereh
AU  - Kalantari T
AD  - Department of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, 
      USA.
FAU - Kamali-Sarvestani, Eskandar
AU  - Kamali-Sarvestani E
FAU - Zhang, Guang-Xian
AU  - Zhang GX
FAU - Safavi, Farinaz
AU  - Safavi F
FAU - Lauretti, Elisabetta
AU  - Lauretti E
FAU - Khedmati, Mohammad-Esmaeil
AU  - Khedmati ME
FAU - Rostami, Abdolmohamad
AU  - Rostami A
LA  - eng
GR  - R01 NS048435/NS/NINDS NIH HHS/United States
PT  - Journal Article
DEP - 20121223
PL  - Netherlands
TA  - Exp Mol Pathol
JT  - Experimental and molecular pathology
JID - 0370711
RN  - 0 (Antigens, Surface)
RN  - 0 (Biomarkers)
RN  - 0 (Cytokines)
RN  - VC2W18DGKR (Thymidine)
SB  - IM
MH  - Animals
MH  - Antigens, Surface/metabolism
MH  - Biomarkers/analysis
MH  - Bone Marrow Cells/*cytology
MH  - *Cell Culture Techniques
MH  - Coculture Techniques
MH  - Cytokines/metabolism
MH  - *Dendritic Cells
MH  - Endocytosis
MH  - Female
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Transgenic
MH  - Spleen
MH  - Stem Cells/*cytology
MH  - Thymidine/metabolism
PMC - PMC3602144
MID - NIHMS443569
EDAT- 2012/12/28 06:00
MHDA- 2013/05/17 06:00
PMCR- 2014/04/01
CRDT- 2012/12/28 06:00
PHST- 2012/12/17 00:00 [received]
PHST- 2012/12/17 00:00 [accepted]
PHST- 2012/12/28 06:00 [entrez]
PHST- 2012/12/28 06:00 [pubmed]
PHST- 2013/05/17 06:00 [medline]
PHST- 2014/04/01 00:00 [pmc-release]
AID - S0014-4800(12)00176-1 [pii]
AID - 10.1016/j.yexmp.2012.12.001 [doi]
PST - ppublish
SO  - Exp Mol Pathol. 2013 Apr;94(2):336-42. doi: 10.1016/j.yexmp.2012.12.001. Epub 
      2012 Dec 23.