8 research outputs found
Isolation and characterization of resident endogenous c-Kitâș cardiac stem cells from the adult mouse and rat heart
This protocol describes the isolation of endogenous c-Kit (also known as CD117)-positive (c-Kitâș), CD45-negative (CD45â») cardiac stem cells (eCSCs) from whole adult mouse and rat hearts. The heart is enzymatically digested via retrograde perfusion of the coronary circulation, resulting in rapid and extensive breakdown of the whole heart. Next, the tissue is mechanically dissociated further and cell fractions are separated by centrifugation. The c-Kitâș CD45â» eCSC population is isolated by magnetic-activated cell sorting technology and purity and cell numbers are assessed by flow cytometry. This process takes âŒ4 h for mouse eCSCs or 4.5 h for rat eCSCs. We also describe how to characterize c-Kitâș CD45â» eCSCs. The c-Kitâș CD45â»eCSCs exhibit the defining characteristics of stem cells: they are self-renewing, clonogenic and multipotent. This protocol also describes how to differentiate eCSCs into three main cardiac lineages: functional, beating cardiomyocytes, smooth muscle, and endothelial cells. These processes take 17-20 d
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Historic overview
The process of learning that neurogenesis actually exists has taken almost a century and has progressed slowly so far. In the late 1800s, scientists worldwide, including the prestigious Spanish researcher, Santiago Ramon y Cajal (1913), maintained that neurogenesis was a process restricted to brain development that ceased after birth. This conclusion was the result of studying the histology of the brain with the techniques of the time, such as Nissl and silver impregnation. Most researchers defined neurons as cells that were characterized by the presence of dendritic arborizations. When dendrites were not well developed, cells were thought to be in the process of differentiation, plastic changes, or the result of a histological artifact. © 2009 Springer-Verlag Berlin Heidelberg
Recommended from our members
Historic overview
The process of learning that neurogenesis actually exists has taken almost a century and has progressed slowly so far. In the late 1800s, scientists worldwide, including the prestigious Spanish researcher, Santiago Ramon y Cajal (1913), maintained that neurogenesis was a process restricted to brain development that ceased after birth. This conclusion was the result of studying the histology of the brain with the techniques of the time, such as Nissl and silver impregnation. Most researchers defined neurons as cells that were characterized by the presence of dendritic arborizations. When dendrites were not well developed, cells were thought to be in the process of differentiation, plastic changes, or the result of a histological artifact. © 2009 Springer-Verlag Berlin Heidelberg