Intravital Microscopy of the Microcirculation in the Mouse Cremaster Muscle for the Analysis of Peripheral Stem Cell Migration

Abstract

In the era of intravascular cell application protocols in the context of regenerative cell therapy, the underlying mechanisms of stem cell migration to nonmarrow tissue have not been completely clarified. We describe here the technique of intravital microscopy applied to the mouse cremaster microcirculation for analysis of peripheral bone marrow stem cell migration in vivo. Intravital microscopy of the M. cremaster has been previously introduced in the field of inflammatory research for direct observation of leucocyte interaction with the vascular endothelium. Since sufficient peripheral stem and progenitor cell migration includes similar initial steps of rolling along and firm adhesion at the endothelial lining it is conceivable to apply the M. cremaster model for the observation and quantification of the interaction of intravasculary administered stem cells with the endothelium. As various chemical components can be selectively applied to the target tissue by simple superfusion techniques, it is possible to establish essential microenvironmental preconditions, for initial stem cell recruitment to take place in a living organism outside the bone marrow.

Actually Seeing What Is Going on – Intravital Microscopy in Tissue Engineering

“…The testicle is either pushed back into the abdominal cavity or removed by orchiectomy. The remaining cremaster muscle is spread over the customized stage and can be accessed for microscopy and interventions ( Bagher and Segal, 2011 ; Reichel et al, 2011 ; Donndorf et al, 2013 ).…”

“…This well-standardized surgical procedure can be a useful tool for visualizing and analyzing capillary perfusion, leukocyte–endothelial interaction, microvascular response to different stimuli and endothelial permeability in a defined environment ( Donndorf et al, 2013 ; Molski et al, 2015 ). It can also be used to study blood cell interactions under influence of different drugs and chemokines ( Reichel et al, 2012 ; Rius and Sanz, 2015 ) as well as ischemia-reperfusion (IR) injury and local effects of systemic conditions ( Molski et al, 2015 ).…”

Intravital Imaging of Hepatic Blood Biliary Barrier in Live Mice

“…Intravital live imaging has emerged as one of the powerful tools to image physiologic processes in real time (Bennewitz, Watkins, & Sundd, 2014; Weigert, Porat‐Shliom, & Amornphimoltham, 2013). It has been routinely applied to many tissues including the brain (De Niz, Nacer, & Frischknecht, 2019; Ricard & Debarbieux, 2014), cremaster (Donndorf et al., 2013), lung (see Current Protocols article: Brzoska, Kaminski, Bennewitz, & Sundd, 2020; Bennewitz et al., 2014; Yipp et al., 2017), liver (Pradhan‐Sundd, Vats, et al., 2018), kidney (Hato, Winfree, & Dagher, 2018; Peti‐Peterdi, Kidokoro, & Riquier‐Brison, 2016), and lymph nodes (Meijer et al., 2017; Miller, Wei, Parker, & Cahalan, 2002). Liver intravital imaging is an emerging field that has been instrumental in understanding three‐dimensional dynamics of liver tissue including immune environment (Liew & Kubes, 2016; Matsumoto, Kikuta, & Ishii, 2018), dynamics of the tumor microenvironment (Babes & Kubes, 2016), barrier function, and pathologic manifestation of liver disease progression (Pradhan‐Sundd, Vats, et al., 2018; Pradhan‐Sundd, Zhou, et al., 2018; Vats et al., 2020).…”