PMID- 27774933
OWN - NLM
STAT- MEDLINE
DCOM- 20170613
LR  - 20171116
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 32
IP  - 11
DP  - 2016 Nov
TI  - [Acquirement of autologous murine cytotoxic T lymphocytes via cryopreservation of 
      lymphocytes].
PG  - 1453-1457
AB  - Objective To evaluate the effects of cryopreservation on the proliferation and 
      killing activity of lymphocytes, and explore a novel protocol of preparing 
      autologous mouse cytotoxic T lymphocytes (CTLs). Methods Mononuclear cells 
      derived from spleen (5.0x10(6)/mL) were cryopreserved in CELLBANKER2 for 6 days 
      and recovered in water bath at 39DegreesCelsius. The fresh lymphocytes and 
      post-cryopreservation lymphocytes were induced by CD3 mAb (100 ng/mL) and 
      recombinant mouse interleukin 2 (rmIL-2, 100 ng/mL) to obtain cytokine-induced 
      killer cells (CIKs). Dendritic cells (DCs) were co-cultured with fresh allogenic 
      lymphocytes and post-cryopreservation autologous lymphocytes to obtain CTLs. The 
      viable cells were counted by trypan blue staining; the percentages of CD3(+)T 
      cells and regulatory T cells (Tregs) were determined by flow cytometry; the 
      levels of supernatant IFN-gamma were detected through ELISA and the cytotoxicity was 
      evaluated by lactate dehydrogenase (LDH) assay. Results The rate of viable 
      lymphocytes declined to 78% after cryopreservation, and there were no significant 
      differences in the percentages of CD3(+)T cells and Tregs between 
      pre-cryopreservation and post-cryopreservation. There were no significant 
      differences in the proliferation of Tregs, the level of IFN-gamma and the 
      cytotoxicity between the fresh CIKs and cryopreservation CIKs, and the similar 
      results were get between the autologous CTLs and allogenic CTLs. Conclusion The 
      autologous CTLs acquired via cryopreservation of lymphocytes is equivalent to the 
      allogenic CTLs with the similar proliferation and killing activity in vitro.
FAU - Wang, Lei
AU  - Wang L
AD  - Zhongnan Hospital, Institute of Liver and Gallbladder Disease, Transplant Center, 
      Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, 
      Wuhan 430071, China.
FAU - Peng, Na
AU  - Peng N
AD  - Zhongnan Hospital, Institute of Liver and Gallbladder Disease, Transplant Center, 
      Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, 
      Wuhan 430071, China.
FAU - Hu, Xiaoyan
AU  - Hu X
AD  - Zhongnan Hospital, Institute of Liver and Gallbladder Disease, Transplant Center, 
      Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, 
      Wuhan 430071, China.
FAU - Liang, Wentao
AU  - Liang W
AD  - Institute of General Surgery, Chinese PLA General Hospital, Beijing 100853, 
      China.
FAU - Liang, Kai
AU  - Liang K
AD  - Institute of General Surgery, Chinese PLA General Hospital, Beijing 100853, 
      China.
FAU - Peng, Guizhu
AU  - Peng G
AD  - Zhongnan Hospital, Institute of Liver and Gallbladder Disease, Transplant Center, 
      Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, 
      Wuhan 430071, China. *Corresponding author, E-mail: pengguizhu@139.com.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (CD3 Complex)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Animals
MH  - CD3 Complex/metabolism
MH  - Cells, Cultured
MH  - Cryopreservation/*methods
MH  - Cytotoxicity, Immunologic
MH  - Dendritic Cells/cytology/metabolism
MH  - Interferon-gamma/metabolism
MH  - Lymphocytes/*cytology/immunology
MH  - Mice
MH  - T-Lymphocytes/cytology/immunology
MH  - T-Lymphocytes, Cytotoxic/*cytology/immunology
EDAT- 2016/10/25 06:00
MHDA- 2017/06/14 06:00
CRDT- 2016/10/25 06:00
PHST- 2016/10/25 06:00 [pubmed]
PHST- 2017/06/14 06:00 [medline]
PHST- 2016/10/25 06:00 [entrez]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Nov;32(11):1453-1457.