Bat-mouse bone marrow chimera: a novel animal model for dissecting the uniqueness of the bat immune system

Yong, Kylie Su Mei; Ng, Justin Han Jia; Her, Zhisheng; Hey, Ying Ying; Tan, Sue Yee; Tan, Wilson Wei Sheng; Irac, Sergio Erdal; Liu, Min; Chan, Xue Ying; Gunawan, Merry; Foo, Randy Jee Hiang; Low, Dolyce Hong Wen; Mendenhall, Ian Hewitt; Chionh, Yok Teng; Dutertre, Charles-Antoine; Chen, Qingfeng; Wang, Lin-Fa

Cell proliferation assay. Mesenteric lymph nodes from bat-mice were harvested, processed into single cell suspension and incubated with Cell Trace Violet (Invitrogen, USA) (0.5 μM final concentration) for 20 minutes at 37 °C with 5% CO 2 . To quench the reaction, 10% FBS was added to the cells and incubated for 5 minutes on ice. The cells were washed twice in PBS supplemented with 2% FBS and resuspended in complete medium, seeded at 1 × 10 5 cells per well in a 96-well flat bottom plate and cultured for 5 days at 37 °C, 5% CO 2 , with either 2 μg/ml SCIenTIfIC REPoRTS | (2018) 8:4726 | DOI:10.1038/s41598-018-22899-1 concanavalin A (ConA) (Sigma), KLH (Biosearch Technologies, USA) or media alone. After 5 days incubation, cells were stained with specific antibodies and analyzed by flow cytometry (as described above).