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Proceedings Paper

Second-harmonic generation microscopy used to evaluate the effect of the dimethyl sulfoxide in the cryopreservation process in collagen fibers of differentiated chondrocytes
Author(s): M. F. Andreoli-Risso; A. S. S. Duarte; T. B. Ribeiro; P. Bordeaux-Rego; A. Luzo; M. O. Baratti; J. Adur; A. A. de Thomaz; V. B. Pelegati; H. F. Carvalho; C. L. Cesar; P. Kharmadayan; F. F. Costa; S. T. Olalla-Saad
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Paper Abstract

Cartilaginous lesions are a significant public health problem and the use of adult stem cells represents a promising therapy for this condition. Cryopreservation confers many advantages for practitioners engaged in cell-based therapies. However, conventional slow freezing has always been associated with damage and mortality due to intracellular ice formation, cryoprotectant toxicity, and dehydration. The aim of this work is to observe the effect of the usual Dimethyl Sulfoxide (DMSO) cryopreservation process on the architecture of the collagen fiber network of chondrogenic cells from mesenchymal stem cells by Second Harmonic Generation (SHG) microscopy. To perform this study we used Mesenchymal Stem Cells (MSC) derived from adipose tissue which presents the capacity to differentiate into other lineages such as osteogenic, adipogenic and chondrogenic lineages. Mesenchymal stem cells obtained after liposuction were isolated digested by collagenase type I and characterization was carried out by differentiation of mesodermic lineages, and flow cytometry using specific markers. The isolated MSCs were cryopreserved by the DMSO technique and the chondrogenic differentiation was carried out using the micromass technique. We then compared the cryopreserved vs non-cryopreserved collagen fibers which are naturally formed during the differentiation process. We observed that noncryopreserved MSCs presented a directional trend in the collagen fibers formed which was absent in the cryopreserved MSCs. We confirmed this trend quantitatively by the aspect ratio obtained by Fast Fourier Transform which was 0.76 for cryopreserved and 0.52 for non-cryopreserved MSCs, a statistical significant difference.

Paper Details

Date Published: 9 February 2012
PDF: 10 pages
Proc. SPIE 8226, Multiphoton Microscopy in the Biomedical Sciences XII, 82263G (9 February 2012); doi: 10.1117/12.909284
Show Author Affiliations
M. F. Andreoli-Risso, Univ. Estadual de Campinas (Brazil)
A. S. S. Duarte, Univ. Estadual de Campinas (Brazil)
T. B. Ribeiro, Univ. Estadual de Campinas (Brazil)
P. Bordeaux-Rego, Univ. Estadual de Campinas (Brazil)
A. Luzo, Univ. Estadual de Campinas (Brazil)
M. O. Baratti, Univ. Estadual de Campinas (Brazil)
J. Adur, Univ. Estadual de Campinas (Brazil)
National Univ. of Entre Rios (Argentina)
A. A. de Thomaz, Univ. Estadual de Campinas (Brazil)
V. B. Pelegati, Univ. Estadual de Campinas (Brazil)
H. F. Carvalho, National Univ. of Entre Rios (Argentina)
C. L. Cesar, Univ. Estadual de Campinas (Brazil)
P. Kharmadayan, Univ. Estadual de Campinas (Brazil)
F. F. Costa, Univ. Estadual de Campinas (Brazil)
S. T. Olalla-Saad, Univ. Estadual de Campinas (Brazil)

Published in SPIE Proceedings Vol. 8226:
Multiphoton Microscopy in the Biomedical Sciences XII
Karsten König, Editor(s)

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