PMID- 34029621
OWN - NLM
STAT- MEDLINE
DCOM- 20211025
LR  - 20240226
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 495
DP  - 2021 Aug
TI  - An optimized method for the induction and purification of mouse bone marrow 
      dendritic cells.
PG  - 113073
LID - S0022-1759(21)00118-6 [pii]
LID - 10.1016/j.jim.2021.113073 [doi]
AB  - Dendritic cells (DCs) play an essential role in the initiation of adaptive immune 
      responses, but they are rare in all organs. The traditional methods used to 
      increase the yield and purity of DCs are the early removal of granulocyte culture 
      medium and the isolation of high-purity DCs by magnetic-activated cell sorting 
      (MACS). This study provides a more rapid and economical optimization method to 
      obtain more high-purity DCs. (i) We harvested 18% more bone marrow (BM) cells by 
      using forceps to crack the epiphysis instead of cutting it with scissors during 
      BM cell extraction. (ii) When the cells in the culture medium that is discarded 
      on day 3 in the traditional method were centrifuged and then added back to the 
      petri dish, the DC yield on day 5 increased by 61%. (iii) On the third day, the 
      addition of fresh medium and the retention of the original medium rather than 
      discarding it increased the number of DCs harvested on the fifth day by 137%. 
      (i-iii) The improved method cost an average of 74% less than the conventional 
      method and yielded the same number and function of cells. (iv) The initial number 
      of BM cells was increased by 15% in 4-week-old mice compared with 8-week-old 
      mice. (v) The Percoll density centrifugation (PDS) method was used to purify DCs 
      on day 6 after induction, and the purity of the DCs was greater than 90%, which 
      showed no significant difference from the MACS method. However, the yield of the 
      PDS method increased by 21%. In addition, the PDS method has a lower cost, with 
      an average purification cost of 4 CNY ($0.58) compared with 648 CNY ($93.25) for 
      MACS, reducing the cost by 99%. Therefore, high-purity and high-yield DCs can be 
      rapidly obtained through a five-step improvement in the process of BM cell 
      extraction, induction and purification.
CI  - Copyright (c) 2021 The Authors. Published by Elsevier B.V. All rights reserved.
FAU - Liu, Ling
AU  - Liu L
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Fan, Shanwen
AU  - Fan S
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Lu, Zhonghua
AU  - Lu Z
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Chen, Zhenxing
AU  - Chen Z
AD  - Department of Gastroenterology, Zhongda Hospital, Southeast University School of 
      Medicine, Nanjing, China.
FAU - Chu, Cuilin
AU  - Chu C
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Liu, Airan
AU  - Liu A
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Xia, Feiping
AU  - Xia F
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Meng, Shanshan
AU  - Meng S
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Guo, Fengmei
AU  - Guo F
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Qiu, Haibo
AU  - Qiu H
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China.
FAU - Yang, Yi
AU  - Yang Y
AD  - Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of 
      Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast 
      University, Nanjing, China. Electronic address: yiyiyang2004@163.com.
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210521
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Biomarkers)
SB  - IM
MH  - *Adaptive Immunity
MH  - Animals
MH  - Biomarkers/metabolism
MH  - Bone Marrow Cells/*immunology/metabolism
MH  - Cell Proliferation
MH  - Cell Separation/economics/*methods
MH  - Cells, Cultured
MH  - Coculture Techniques
MH  - Cost Savings
MH  - Cost-Benefit Analysis
MH  - Dendritic Cells/*immunology/metabolism
MH  - Lymphocyte Activation
MH  - Male
MH  - Mice, Inbred C57BL
MH  - Phagocytosis
MH  - Phenotype
MH  - Time Factors
MH  - Workflow
MH  - Mice
OTO - NOTNLM
OT  - Cell culture
OT  - Dendritic cells
OT  - Magnetic-activated cell sorting
OT  - Percoll
OT  - Purity
EDAT- 2021/05/25 06:00
MHDA- 2021/10/26 06:00
CRDT- 2021/05/24 20:23
PHST- 2020/12/06 00:00 [received]
PHST- 2021/05/01 00:00 [revised]
PHST- 2021/05/13 00:00 [accepted]
PHST- 2021/05/25 06:00 [pubmed]
PHST- 2021/10/26 06:00 [medline]
PHST- 2021/05/24 20:23 [entrez]
AID - S0022-1759(21)00118-6 [pii]
AID - 10.1016/j.jim.2021.113073 [doi]
PST - ppublish
SO  - J Immunol Methods. 2021 Aug;495:113073. doi: 10.1016/j.jim.2021.113073. Epub 2021 
      May 21.