期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 147, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/59342
关键词
Developmental Biology; Issue 147; stem cell; endothelial progenitor; vasculogenesis; extracellular matrix; collagen; computational analysis; vessel networks
资金
- American Heart Association [15SDG25740035]
- National Institute of Biomedical Imaging and Bioengineering (NIBIB) of the National Institutes of Health [EB007507]
- Alliance for Regenerative Rehabilitation Research & Training (AR3T) [1 P2C HD086843-01]
Induced pluripotent stem cells (iPSCs) are a patient-specific, proliferative cell source that can differentiate into any somatic cell type. Bipotent endothelial progenitors (EPs), which can differentiate into the cell types necessary to assemble mature, functional vasculature, have been derived from both embryonic and induced pluripotent stem cells. However, these cells have not been rigorously evaluated in three-dimensional environments, and a quantitative measure of their vasculogenic potential remains elusive. Here, the generation and isolation of iPSC-EPs via fluorescent-activated cell sorting are first outlined, followed by a description of the encapsulation and culture of iPSC-EPs in collagen hydrogels. This extracellular matrix (ECM)-mimicking microenvironment encourages a robust vasculogenic response; vascular networks form after a week of culture. The creation of a computational pipeline that utilizes open-source software to quantify this vasculogenic response is delineated. This pipeline is specifically designed to preserve the 3D architecture of the capillary plexus to robustly identify the number of branches, branching points, and the total network length with minimal user input.
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