Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures.
Cell-in-cell structures (CICs), characterized by the presence of one or more viable cells inside another one, were recently found important player in development, immune homeostasis and tumorigenesis etc. Incompatible with ever-increasing interests on this unique phenomenon, reliable methods available for high throughput quantification and systemic investigation are lacking. Here, we report a flow cytometry-based method for rapid analysis and sorting of heterotypic CICs formed between lymphocytes and tumor cells. In this method, cells were labeled with fluorescent dyes for fluorescence-activated cell sorting (FACS) by flow cytometry, conditions for reducing cell doublets were optimized such that high purity (>95%) of CICs could be achieved. By taking advantage of this method, we analyzed CICs formation between different cell pairs, and found that factors from both internalized effector cells and engulfing target cells affect heterotypic CICs formation. Thus, flow cytometry-based FACS analysis would serve as a high throughput method to promote systemic researches on CICs.
1] Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-Sen University, 58 Zhongshan er Road, Guangzhou, Guangdong 510080, P. R. China [2] Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China [3] The Institute of Life Sciences, the Key Laboratory of Normal Aging &Geriatric, the State Key Laboratory of Kidney, the Chinese PLA General Hospital, Beijing 100853, P. R. China.
Meifang
M
He
1] Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China [2] Department of Oncology, Beijing Shijitan Hospital of Capital Medical University, 10 TIEYI Road, Beijing 100038, P. R. China.
Hongyan
H
Huang
1] Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China [2] Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, P. R. China.
Manna
M
Wang
Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China.
Ang
A
Chen
1] Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China [2] Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, P. R. China.
Xiangkai
X
Ning
Department of Stomatology, Affiliated Hospital of Academy of Military Medical Science, 8 Dongda Street, Beijing 100071, P. R. China.
Kaitao
K
Yu
Department of Stomatology, Affiliated Hospital of Academy of Military Medical Science, 8 Dongda Street, Beijing 100071, P. R. China.
Qihong
Q
Li
Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-Sen University, 58 Zhongshan er Road, Guangzhou, Guangdong 510080, P. R. China.
Wen
W
Li
Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou 510515, P. R. China.
Li
L
Ma
Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China.
Zhaolie
Z
Chen
The Institute of Life Sciences, the Key Laboratory of Normal Aging &Geriatric, the State Key Laboratory of Kidney, the Chinese PLA General Hospital, Beijing 100853, P. R. China.
Xiaoning
X
Wang
1] Laboratory of Cell Engineering, Institute of Biotechnology, 20 Dongda Street, Beijing 100071, P. R. China [2] National Key Laboratory of Medical Immunology and Institute of Immunology, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, P. R. China.
Qiang
Q
Sun
D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D061986:MCF-7 Cells
D005456:Fluorescent Dyes