Open AccessConfocal scanning of intervertebral disc cells in 3D : Inside alginate beads and in native microenvironment
Author(s) -
Hernandez Paula A.,
Jacobsen Timothy D.,
Barati Zahra,
Chahine Nadeen O.
Publication year - 2020
Publication title -
jor spine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.125
0ISSN - 2572-1143
DOI - 10.1002/jsp2.1106
Subject(s) - extracellular matrix , microbiology and biotechnology , confocal microscopy , phenotype , confocal , cytoskeleton , cell , tissue engineering , chemistry , staining , nucleus , biophysics , biology , biochemistry , gene , genetics , geometry , mathematics
The interaction between cells and their extracellular matrix (ECM) is crucial to maintain both tissue and cellular homeostasis. Indeed, cell phenotype is significantly affected by the 3D microenvironment. Although highly convenient, isolating cells from the intervertebral disc (IVD) and growing them in 2D on plastic or glass substrates, causes them to rapidly lose their phenotype and consequently alter their gene and protein expression. While characterization of cells in their native or simulated 3D environment is preferred, such approaches are complexed by limitations in phenotypic readouts. In the current article, we describe a detailed protocol to study nucleus pulposus cells in 3D—embedded in alginate as a permeable cell‐staining reservoir, as well as adaptation for cell staining and imaging in their native ECM. This method allows for detection of phenotypical and cytoskeletal changes in cells within native tissue or 3D alginate beads using confocal microscopy, without the need for histological processing.