TY - JOUR
T1 - Culturing mammalian cells in three-dimensional peptide scaffolds
AU - Betriu, Nausika
AU - Recha-Sancho, Lourdes
AU - Semino, Carlos E.
N1 - Funding Information:
The research performed by the authors was supported in part by grants from the European Union Seventh Framework Programme (FP7/2007-2013) under Grant Agreement No. 229239, and from the AO Foundation, Exploratory Research Collaborative Research Program Acute Cartilage Injury/lesion/Defect (CRP ACI) under the project Bioactive and Biomimetic Scaffolds for Cartilage Regeneration (BIOCART).
Publisher Copyright:
© 2018, Journal of Visualized Experiments. All rights reserved.
PY - 2018/6/13
Y1 - 2018/6/13
N2 - A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an environment that closely mimics the structural features of non-polarized tissue. Furthermore, the particular intrinsic nanofiber structure of the scaffold makes it transparent to visual light, which allows for easy visualization of the sample under microscopy. This advantage was largely used to study cell migration, organization, proliferation, and differentiation and thus any development of their particular cellular function by staining with specific dyes or probes. Furthermore, in this work, we describe the good performance of this system to easily study the redifferentiation of expanded human articular chondrocytes into cartilaginous tissue. Cells were encapsulated into self-assembling peptide scaffolds and cultured under specific conditions to promote chondrogenesis. Three-dimensional cultures showed good viability during the 4 weeks of the experiment. As expected, samples cultured with chondrogenic inducers (compared to non-induced controls) stained strongly positive for toluidine blue (which stains glycosaminoglycans (GAGs) that are highly present in cartilage extracellular matrix) and expressed specific molecular markers, including collagen type I, II and X, according to Western Blot analysis. This protocol is easy to perform and can be used at research laboratories, industries and for educational purposes in laboratory courses.
AB - A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an environment that closely mimics the structural features of non-polarized tissue. Furthermore, the particular intrinsic nanofiber structure of the scaffold makes it transparent to visual light, which allows for easy visualization of the sample under microscopy. This advantage was largely used to study cell migration, organization, proliferation, and differentiation and thus any development of their particular cellular function by staining with specific dyes or probes. Furthermore, in this work, we describe the good performance of this system to easily study the redifferentiation of expanded human articular chondrocytes into cartilaginous tissue. Cells were encapsulated into self-assembling peptide scaffolds and cultured under specific conditions to promote chondrogenesis. Three-dimensional cultures showed good viability during the 4 weeks of the experiment. As expected, samples cultured with chondrogenic inducers (compared to non-induced controls) stained strongly positive for toluidine blue (which stains glycosaminoglycans (GAGs) that are highly present in cartilage extracellular matrix) and expressed specific molecular markers, including collagen type I, II and X, according to Western Blot analysis. This protocol is easy to perform and can be used at research laboratories, industries and for educational purposes in laboratory courses.
KW - 3D-culture
KW - Articular chondrocytes
KW - Bioengineering
KW - Differentiation
KW - Issue 136
KW - Mammal cells
KW - Nanobiomaterials
KW - Nanomaterials
KW - Self-assembling peptide scaffolds
UR - http://www.scopus.com/inward/record.url?scp=85049847015&partnerID=8YFLogxK
UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=pure_univeritat_ramon_llull&SrcAuth=WosAPI&KeyUT=WOS:000444752100039&DestLinkType=FullRecord&DestApp=WOS_CPL
U2 - 10.3791/57259
DO - 10.3791/57259
M3 - Article
C2 - 29985312
AN - SCOPUS:85049847015
SN - 1940-087X
VL - 2018
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 136
M1 - e57259
ER -