Cryopreservation of Murine Spleen Cells

Spleen cells were harvested from Balb/c mice and red blood cells were dissolved with red blood cell lysis buffer (Chinese Academy of Medical Sciences, Beijing, China). After repeated centrifugation and washing, the remaining mononuclear cells were re-suspended at a density of 1 × 10⁷/ml in a cryogenic vial by CELLBANKER 2 (Nippon Zenyaku Kogyo Co., Ltd, Koriyama, Japan). The cells were cryopreserved as follows: 4°C for 3 h, -20°C for 6 h, and -80°C for 2 days stored in liquid nitrogen, and then recovered at 39°C for 5 min, as previously described [20 (link)].

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Wang L., Liang W., Peng N., Hu X., Xu Y, & Liu Z. (2017). The synergistic antitumor effect of arsenic trioxide combined with cytotoxic T cells in pulmonary metastasis model of colon cancer. Oncotarget, 8(65), 109609-109618.

Publication 2017

CELLBANKER 2

Balb c mice Buffer Cells Centrifugation Chinese Nitrogen Red blood cell Spleen

Corresponding Organization :

Other organizations : Fujian Medical University, Dalian Medical University, First Affiliated Hospital of Dalian Medical University, Mengchao Hepatobiliary Hospital, Chinese PLA General Hospital, Zhongnan Hospital of Wuhan University, Wuhan University

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Variable analysis

independent variables

Harvesting spleen cells from Balb/c mice
Dissolving red blood cells with red blood cell lysis buffer
Repeated centrifugation and washing of the remaining mononuclear cells
Re-suspending the mononuclear cells at a density of 1 × 10^7/ml in a cryogenic vial by CELLBANKER 2
Cryopreservation process (4°C for 3 h, -20°C for 6 h, and -80°C for 2 days stored in liquid nitrogen)
Recovering the cells at 39°C for 5 min

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned in the protocol.

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