Cell kinetics, DNA integrity, differentiation, and lipid fingerprinting analysis of rabbit adipose-derived stem cells

Cell kinetics, DNA integrity, differentiation, and lipid fingerprinting analysis of rabbit adipose-derived stem cells

Author Sa Barretto, Leticia Siqueira de Autor UNIFESP Google Scholar
Lessio, Camila Autor UNIFESP Google Scholar
Sawaki e Nakamura, Ahy Natally Autor UNIFESP Google Scholar
Lo Turco, Edson Guimaraes Autor UNIFESP Google Scholar
Silva, Camila Gonzaga da Autor UNIFESP Google Scholar
Zambon, Joao Paulo Autor UNIFESP Google Scholar
Gozzo, Fabio Cesar Google Scholar
Pilau, Eduardo Jorge Google Scholar
Almeida, Fernando Goncalves de Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Universidade Estadual de Campinas (UNICAMP)
Abstract Human adipose tissue has been described as a potential alternative reservoir for stem cells. Although studies have been performed in rabbits using autologous adipose-derived stem cells (ADSC), these cells have not been well characterized. the primary objectives of this study were to demonstrate the presence of adipose-derived stem cells isolated from rabbit inguinal fat pads and to characterize them through osteogenic and adipogenic in vitro differentiation and lipid fingerprinting analysis. the secondary objective was to evaluate cell behavior through growth kinetics, cell viability, and DNA integrity. Rabbit ADSCs were isolated to determine the in vitro growth kinetics and cell viability. DNA integrity was assessed by an alkaline Comet assay in passages 0 and 5. the osteogenic differentiation was evaluated by Von Kossa, and Alizarin Red S staining and adipogenic differentiation were assessed by Oil Red O staining. Lipid fingerprinting analyses of control, adipogenic, and osteogenic differentiated cells were performed by MALDI-TOF/MS. We demonstrate that rabbit ADSC have a constant growth rate at the early passages, with increased DNA fragmentation at or after passage 5. Rabbit ADSC viability was similar in passages 2 and 5 (90.7% and 86.6%, respectively), but there was a tendency to decreased cellular growth rate after passage 3. the ADSC were characterized by the expression of surface markers such as CD29 (67.4%) and CD44 (89.4%), using CD 45 (0.77%) as a negative control. ADSC from rabbits were successfully isolated form the inguinal region. These cells were capable to differentiate into osteogenic and adipogenic tissue when they were placed in inductive media. After each passage, there was a trend towards decreased cell growth. On the other hand, DNA fragmentation increased at each passage. ADSC had a different lipid profile when placed in control, adipogenic, or osteogenic media.
Keywords Adipose-derived stem cells
Rabbit
DNA integrity
Lipid fingerprinting analysis
Animal model
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Grant number FAPESP: 06/57479-2
Date 2014-10-01
Published in In Vitro Cellular & Developmental Biology-animal. New York: Springer, v. 50, n. 9, p. 831-839, 2014.
ISSN 1071-2690 (Sherpa/Romeo, impact factor)
Publisher Springer
Extent 831-839
Origin http://dx.doi.org/10.1007/s11626-014-9782-x
Access rights Closed access
Type Article
Web of Science ID WOS:000344389900008
URI http://repositorio.unifesp.br/handle/11600/38273

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