title: Human Adipose-Derived Mesenchymal Stem Cells Cryopreservation and Thawing Decrease α4-Integrin Expression.
abstract: Aim. The effects of cryopreservation on adipose tissue-derived mesenchymal stem cells are not clearly documented, as there is a growing body of evidence about the importance of adipose-derived mesenchymal stem cells for regenerative therapies. The aim of this study was to analyze human adipose tissue-derived mesenchymal stem cells phenotypic expression (CD34, CD45, CD73, CD90, CD105, and CD49d), colony forming unit ability, viability, and differentiation potential before and after cryopreservation. Materials and Methods. 12 samples of the adipose tissue were collected from a healthy donor using the liposuction technique. The cell isolation was performed by enzymatic digestion and then the cells were cultured up to passage 2. Before and after cryopreservation the immunophenotype, cellular viability analysis by flow cytometer, colony forming units ability, differentiation potential into adipocytes and osteoblasts as demonstrated by Oil Red O and Alizarin Red staining, respectively. Results. The immunophenotypic markers expression was largely preserved, and their multipotency was maintained. However, after cryopreservation, the cells decreased α4-integrin expression (CD49d), cell viability, and number of colony forming units. Conclusions. These findings suggest that ADMSC transplanted after cryopreservation might compromise the retention of transplanted cells in the host tissue. Therefore, further studies are warranted to standardize protocols related to cryopreservation to attain full benefits of stem cell therapy.
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil; Bioprocess Engineering and Biotechnology Department, Federal University of Paraná, Avenida Coronel Francisco Heráclito dos Santos 210, 81531-970 Curitiba, PR, Brazil.
authors.forename: Ana Carolina
authors.initials: AC
authors.lastname: Irioda
authors.affiliation: Hospital Santé Curitiba, Plastic Surgery Clinic, Rua XV de Novembro 2913, 80045-340 Curitiba, PR, Brazil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Cassilha
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Larissa
authors.initials: L
authors.lastname: Zocche
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Julio Cesar
authors.initials: JC
authors.lastname: Francisco
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Ricardo Correa
authors.initials: RC
authors.lastname: Cunha
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Priscila Elias
authors.initials: PE
authors.lastname: Ferreira
authors.affiliation: Experimental Laboratory of Institute of Biological and Health Sciences of Pontifical Catholic University of Paraná (PUCPR), Rua Imaculada Conceição 1155, 80215-901 Curitiba, PR, Brazil.
authors.forename: Luiz Cesar
authors.initials: LC
authors.lastname: Guarita-Souza
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Reginaldo Justino
authors.initials: RJ
authors.lastname: Ferreira
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Bassam Felipe
authors.initials: BF
authors.lastname: Mogharbel
authors.affiliation: Center for Translational Medicine, Temple University School of Medicine, Philadelphia, PA 19140, USA.
authors.forename: Venkata Naga Srikanth
authors.initials: VN
authors.lastname: Garikipati
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil.
authors.forename: Daiany
authors.initials: D
authors.lastname: Souza
authors.affiliation: Flow Cytometric Analysis Laboratory, Clinical Hospital of Federal University of Paraná, Rua Padre Camargo 280, Alto da Glória, 80060-240 Curitiba, PR, Brazil.
authors.forename: Mirian Perlingeiro
authors.initials: MP
authors.lastname: Beltrame
authors.affiliation: Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil; Bioprocess Engineering and Biotechnology Department, Federal University of Paraná, Avenida Coronel Francisco Heráclito dos Santos 210, 81531-970 Curitiba, PR, Brazil.
authors.forename: Katherine Athayde Teixeira
authors.initials: KA
authors.lastname: de Carvalho