PMID- 37300611
OWN - NLM
STAT- MEDLINE
DCOM- 20230612
LR  - 20231117
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2679
DP  - 2023
TI  - Functional Interrogation of Ca(2+) Signals in Human Cancer Cells In Vitro and Ex 
      Vivo by Fluorescent Microscopy and Molecular Tools.
PG  - 95-125
LID - 10.1007/978-1-0716-3271-0_7 [doi]
AB  - Genetically encoded calcium indicators (GECIs) and high-resolution confocal 
      microscopy enable dynamic visualization of calcium signals in cells and tissues. 
      Two-dimensional and 3D biocompatible materials mimic the mechanical 
      microenvironments of tumor and healthy tissues in a programmable manner. Cancer 
      xenograft models and ex vivo functional imaging of tumor slices reveal 
      physiologically relevant functions of calcium dynamics in tumors at different 
      progression stages. Integration of these powerful techniques allows us to 
      quantify, diagnose, model, and understand cancer pathobiology. Here, we describe 
      detailed materials and methods used to establish this integrated interrogation 
      platform, from generating transduced cancer cell lines that stably express CaViar 
      (GCaMP5G + QuasAr2) to in vitro and ex vivo calcium imaging of the cells in 2D/3D 
      hydrogels and tumor tissues. These tools open the possibility for detailed 
      explorations of mechano-electro-chemical network dynamics in living systems.
CI  - (c) 2023. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Liang, Chenyu
AU  - Liang C
AD  - Department of Mechanical & Aerospace Engineering, Herbert Wertheim College of 
      Engineering, University of Florida, Gainesville, FL, USA.
FAU - Huang, Miao
AU  - Huang M
AD  - Department of Mechanical & Aerospace Engineering, Herbert Wertheim College of 
      Engineering, University of Florida, Gainesville, FL, USA.
FAU - Tanaka, Mai
AU  - Tanaka M
AD  - Department of Radiation Oncology, College of Medicine, University of Florida, 
      Gainesville, FL, USA.
FAU - Lightsey, Suzanne
AU  - Lightsey S
AD  - Department of Biomedical Engineering, Herbert Wertheim College of Engineering, 
      University of Florida, Gainesville, FL, USA.
FAU - Temples, Madison
AU  - Temples M
AD  - Department of Biomedical Engineering, Herbert Wertheim College of Engineering, 
      University of Florida, Gainesville, FL, USA.
FAU - Lepler, Sharon E
AU  - Lepler SE
AD  - Department of Radiation Oncology, College of Medicine, University of Florida, 
      Gainesville, FL, USA.
FAU - Sheng, Peike
AU  - Sheng P
AD  - Department of Biochemistry and Molecular Biology, College of Medicine, University 
      of Florida, Gainesville, FL, USA.
FAU - Mann, William P
AU  - Mann WP
AD  - Department of Mechanical & Aerospace Engineering, Herbert Wertheim College of 
      Engineering, University of Florida, Gainesville, FL, USA.
FAU - Widener, Adrienne E
AU  - Widener AE
AD  - Department of Biomedical Engineering, Herbert Wertheim College of Engineering, 
      University of Florida, Gainesville, FL, USA.
FAU - Siemann, Dietmar W
AU  - Siemann DW
AD  - Department of Radiation Oncology, College of Medicine, University of Florida, 
      Gainesville, FL, USA.
FAU - Sharma, Blanka
AU  - Sharma B
AD  - Department of Biomedical Engineering, Herbert Wertheim College of Engineering, 
      University of Florida, Gainesville, FL, USA.
FAU - Xie, Mingyi
AU  - Xie M
AD  - Department of Biochemistry and Molecular Biology, College of Medicine, University 
      of Florida, Gainesville, FL, USA.
FAU - Dai, Yao
AU  - Dai Y
AD  - Department of Radiation Oncology, College of Medicine, University of Florida, 
      Gainesville, FL, USA.
FAU - Phelps, Edward
AU  - Phelps E
AD  - Department of Biomedical Engineering, Herbert Wertheim College of Engineering, 
      University of Florida, Gainesville, FL, USA.
FAU - Zeng, Bo
AU  - Zeng B
AD  - Institute of Cardiovascular Research, Southwest Medical University, Luzhou, 
      China. zengbo@swmu.edu.cn.
FAU - Tang, Xin
AU  - Tang X
AD  - Department of Mechanical & Aerospace Engineering, Herbert Wertheim College of 
      Engineering, University of Florida, Gainesville, FL, USA. xin.tang@ufl.edu.
LA  - eng
GR  - R35 GM128753/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - SY7Q814VUP (Calcium)
RN  - 0 (Indicators and Reagents)
RN  - 0 (Coloring Agents)
SB  - IM
MH  - Humans
MH  - *Calcium/metabolism
MH  - Cell Line
MH  - Indicators and Reagents
MH  - Coloring Agents
MH  - Microscopy, Fluorescence/methods
MH  - *Neoplasms/genetics
MH  - Calcium Signaling/physiology
MH  - Tumor Microenvironment
OTO - NOTNLM
OT  - 2D and 3D hydrogels
OT  - CRISPR/Cas9 gene editing
OT  - Cancer xenograft model
OT  - Genetically encoded calcium indicator (GECIs)
OT  - In vitro and ex vivo calcium imaging
OT  - Mechanobiology
OT  - Optogenetics
OT  - Retrovirus and lentivirus transduction
EDAT- 2023/06/10 23:42
MHDA- 2023/06/12 06:42
CRDT- 2023/06/10 11:07
PHST- 2023/06/12 06:42 [medline]
PHST- 2023/06/10 23:42 [pubmed]
PHST- 2023/06/10 11:07 [entrez]
AID - 10.1007/978-1-0716-3271-0_7 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2023;2679:95-125. doi: 10.1007/978-1-0716-3271-0_7.