An Experimental Model for Simultaneous Study of Migration of Cell Fragments, Single Cells, and Cell Sheets.
Recent studies have demonstrated distinctive motility and responses to extracellular cues of cells in isolation, cells collectively in groups, and cell fragments. Here we provide a protocol for generating cell sheets, isolated cells, and cell fragments of keratocytes from zebrafish scales. The protocol starts with a comprehensive fish preparation, followed by critical steps for scale processing and subsequent cell sheet generation, single cell isolation, and cell fragment induction, which can be accomplished in just 3 days including a 36-48 h incubation time. Compared to other approaches that usually produce single cells only or together with either fragments or cell groups, this facile and reliable methodology allows generation of all three motile forms simultaneously. With the powerful genetics in zebrafish our model system offers a useful tool for comparison of the mechanisms by which cell sheets, single cells, and cell fragments respond to extracellular stimuli.
Department of Dermatology, School of Medicine, University of California-Davis, Sacramento, CA, USA.
Yao-Hui
YH
Sun
Department of Dermatology, School of Medicine, University of California-Davis, Sacramento, CA, USA.|Department of Radiology and Biomedical Imaging, University of California-San Francisco, San Francisco, CA, USA.
Yuxin
Y
Sun
Department of Dermatology, School of Medicine, University of California-Davis, Sacramento, CA, USA.|Bioelectromagnetics Laboratory, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
Kan
K
Zhu
Department of Molecular and Cellular Biology, School of Medicine, University of California-Davis, Sacramento, CA, USA.
Bruce W
BW
Draper
Bioelectromagnetics Laboratory, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
Qunli
Q
Zeng
Courant Institute and Department of Biology, New York University, New York, NY, USA.
Alex
A
Mogilner
Department of Dermatology, School of Medicine, University of California-Davis, Sacramento, CA, USA. minzhao@ucdavis.edu.|Department of Ophthalmology, School of Medicine, University of California-Davis, Sacramento, CA, USA. minzhao@ucdavis.edu.
Min
M
Zhao
D000818:Animals; D018929:Cell Culture Techniques; D002465:Cell Movement; D002478:Cells, Cultured; D005260:Female; D008297:Male; D015027:Zebrafish
Cell fragment; Cell migration; Collective cell migration; Electric fields; Electrotaxis; Galvanotaxis; Zebrafish