TY - JOUR
T1 - Nanoscopic Characterization of Cell Migration under Flow Using Optical and Electron Microscopy
AU - Alghamdi, Abdullah
AU - Tamra, Amar
AU - Rakhmatulina, Aigerim
AU - Nozue, Shuho
AU - Al-Amoodi, Asma S.
AU - Aldehaiman, Mansour M.
AU - Isaioglou, Ioannis
AU - Merzaban, Jasmeen S.
AU - Habuchi, Satoshi
N1 - Funding Information:
The authors thank Umme Habiba for purifying an E-selectin blocking antibody anti-E-selectin (H18-7) from hybridoma mouse H18/7 (ATCC: HB-11684). The research reported in this publication was supported by the King Abdullah University of Science and Technology (KAUST).
Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.
PY - 2022
Y1 - 2022
N2 - Hematopoietic stem/progenitor cell (HSPC) and leukemic cell homing is an important biological phenomenon that takes place through essential interactions with adhesion molecules on an endothelial cell layer. The homing process of HSPCs begins with the tethering and rolling of the cells on the endothelial layer, which is achieved by the interaction between selectins on the endothelium to the ligands on HSPC/leukemic cells under shear stress of the blood flow. Although many studies have been based on in vitro conditions of the cells rolling over recombinant proteins, significant challenges remain when imaging HSPC/leukemic cells on the endothelium, a necessity when considering characterizing cell-to-cell interaction and rolling dynamics during cell migration. Here, we report a new methodology that enables imaging of stem-cell-intrinsic spatiotemporal details during its migration on an endothelium-like cell monolayer. We developed optimized protocols that preserve transiently appearing structures on HSPCs/leukemic cells during its rolling under shear stress for fluorescence and scanning electron microscopy characterization. Our new experimental platform is closer to in vivo conditions and will contribute to indepth understanding of stem-cell behavior during its migration and cell-to-cell interaction during the process of homing.
AB - Hematopoietic stem/progenitor cell (HSPC) and leukemic cell homing is an important biological phenomenon that takes place through essential interactions with adhesion molecules on an endothelial cell layer. The homing process of HSPCs begins with the tethering and rolling of the cells on the endothelial layer, which is achieved by the interaction between selectins on the endothelium to the ligands on HSPC/leukemic cells under shear stress of the blood flow. Although many studies have been based on in vitro conditions of the cells rolling over recombinant proteins, significant challenges remain when imaging HSPC/leukemic cells on the endothelium, a necessity when considering characterizing cell-to-cell interaction and rolling dynamics during cell migration. Here, we report a new methodology that enables imaging of stem-cell-intrinsic spatiotemporal details during its migration on an endothelium-like cell monolayer. We developed optimized protocols that preserve transiently appearing structures on HSPCs/leukemic cells during its rolling under shear stress for fluorescence and scanning electron microscopy characterization. Our new experimental platform is closer to in vivo conditions and will contribute to indepth understanding of stem-cell behavior during its migration and cell-to-cell interaction during the process of homing.
UR - http://www.scopus.com/inward/record.url?scp=85146259607&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.2c04222
DO - 10.1021/acs.analchem.2c04222
M3 - Article
C2 - 36627105
AN - SCOPUS:85146259607
SN - 0003-2700
VL - 95
SP - 1958
EP - 1966
JO - Analytical chemistry
JF - Analytical chemistry
IS - 3
ER -