Fabiana Zanata1, Annie Bowles, Trivia Frazier, J Lowry Curley, Bruce A Bunnell, Xiying Wu, James Wade, Ram Devireddy, Jeffrey M Gimble, Lydia Masako Ferreira. 1. São Paulo and Brasilia, Brazil; and New Orleans, Covington, and Baton Rouge, La. From the Translational Surgery Graduate Program and Plastic Surgery Division, Federal University of São Paulo; the Center for Stem Cell Research & Regenerative Medicine, Tulane University; the Department of Cell and Molecular Biology, Tulane University School of Science and Engineering; the Physics Department, Dillard University; La Cell LLC; the Departments of Pharmacology, Medicine, Structural and Cellular Biology, and Surgery, Tulane University School of Medicine; the Tulane National Primate Research Center; Plastic Surgery Associates; the Department of Mechanical Engineering, Louisiana State University; and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.
Abstract
BACKGROUND: Adipose tissue is a source of adipose-derived stromal/stem cells for tissue engineering and reconstruction and a tissue source for fat grafts. Although liposuction is a simple procedure for the harvest of adipose tissue, the repetition of this surgical intervention can cause adverse effects to the patient and can be a limiting factor for immediate use. Cryopreservation can avoid the morbidity associated with repetitive liposuction, allowing the use of stored tissue after the initial harvest procedure. This article focuses on the characterization of fresh and cryopreserved human adipose tissue. METHODS: Lipoaspirates from eight donors were processed as fresh adipose tissue or cryopreserved for 4 to 6 weeks. Fresh and cryopreserved tissues were collagenase digested and the stromal vascular fraction cells were characterized immediately or cryopreserved. Characterization was based on stromal vascular fraction cell proliferation and immunophenotype. In vivo fat grafting was performed in C57BL/6 green fluorescent protein mice to analyze morphology of the tissue and its adiposity using confocal microscopy, histochemical staining (i.e., hematoxylin and eosin and Masson trichrome), and immunohistochemistry (i.e., green fluorescent protein, perilipin, and CD31). RESULTS: Although tissue and stromal vascular fraction cell cryopreservation reduced the total cell yield, the remaining viable cells retained their adhesive and proliferative properties. The stromal vascular fraction cell immunophenotype showed a significant reduction in the hematopoietic surface markers and increased expression of stromal and adipogenic markers following cryopreservation. In vivo cryopreserved fat grafts showed morphology similar to that of freshly implanted fat grafts. CONCLUSION: In this study, the authors demonstrated that cryopreserved adipose tissue is a potential source of stromal vascular fraction cells and a suitable source for fat grafts.
BACKGROUND: Adipose tissue is a source of adipose-derived stromal/stem cells for tissue engineering and reconstruction and a tissue source for fat grafts. Although liposuction is a simple procedure for the harvest of adipose tissue, the repetition of this surgical intervention can cause adverse effects to the patient and can be a limiting factor for immediate use. Cryopreservation can avoid the morbidity associated with repetitive liposuction, allowing the use of stored tissue after the initial harvest procedure. This article focuses on the characterization of fresh and cryopreserved human adipose tissue. METHODS: Lipoaspirates from eight donors were processed as fresh adipose tissue or cryopreserved for 4 to 6 weeks. Fresh and cryopreserved tissues were collagenase digested and the stromal vascular fraction cells were characterized immediately or cryopreserved. Characterization was based on stromal vascular fraction cell proliferation and immunophenotype. In vivo fat grafting was performed in C57BL/6 green fluorescent protein mice to analyze morphology of the tissue and its adiposity using confocal microscopy, histochemical staining (i.e., hematoxylin and eosin and Masson trichrome), and immunohistochemistry (i.e., green fluorescent protein, perilipin, and CD31). RESULTS: Although tissue and stromal vascular fraction cell cryopreservation reduced the total cell yield, the remaining viable cells retained their adhesive and proliferative properties. The stromal vascular fraction cell immunophenotype showed a significant reduction in the hematopoietic surface markers and increased expression of stromal and adipogenic markers following cryopreservation. In vivo cryopreserved fat grafts showed morphology similar to that of freshly implanted fat grafts. CONCLUSION: In this study, the authors demonstrated that cryopreserved adipose tissue is a potential source of stromal vascular fraction cells and a suitable source for fat grafts.
Authors: Trivia Frazier; Christopher Williams; Michael Henderson; Tamika Duplessis; Emma Rogers; Xiying Wu; Katie Hamel; Elizabeth C Martin; Omair Mohiuddin; Shahensha Shaik; Ram Devireddy; Brian G Rowan; Daniel J Hayes; Jeffrey M Gimble Journal: Tissue Eng Part A Date: 2021-03-10 Impact factor: 3.845
Authors: Ekaterina Semenova; Mariusz P Grudniak; Katarzyna Bocian; Magdalena Chroscinska-Krawczyk; Marzena Trochonowicz; Igor M Stepaniec; Magdalena Murzyn; Ilona Szablowska-Gadomska; Dariusz Boruczkowski; Tomasz Oldak; Eugeniusz K Machaj Journal: Front Bioeng Biotechnol Date: 2021-11-30