A rapid sonication based method for preparation of stromal vascular fraction and mesenchymal stem cells from fat tissue

Abstract

Introduction: Much attention has been paid to the idea of cell therapy using stem cells from different sources of the body. Fat-derived stem cells that are called adipose derived stem cells (ADSCs) from stromal vascular fraction (SVF) are the subject of many studies in several cell therapy clinical trials. Despite production of some GMP-grade enzymes to isolate SVF for clinical trials, there are critical conditions like inconsistency in lot-to-lot enzyme activity, endotoxin residues, other protease activities and cleavage of some cell surface markers which significantly narrow the options. So we decided to develop a new method via sonication cavitation to homogenize fat tissue and disrupt partially adipose cells to obtain SVF and finally ADSCs at a minimum of time and expenses. Methods: The fat tissue was chopped in a sterile condition by a blender mixer and then sonicated for 2 s before centrifugation. The next steps were performed as the regular methods of SVF harvesting, and then it was characterized using flow cytometry. Results: Analysis of the surface markers of the cells revealed similar sets of surface antigens. The cells showed slightly high expression of CD34, CD73 and CD105. The differentiation capacity of these cells indicates that multipotent properties of the cells are not compromised after sonication. But we had the less osteogenic potential of cells when compared with the enzymatic method. Conclusion: The current protocol based on the sonication-mediated cavitation is a rapid, safe and cost-effective method, which is proposed for isolation of SVF and of course ADSCs cultures in a large scale for the clinical trials or therapeutic purposes.

Zinc sulfate contributes to promote telomere length extension via increasing telomerase gene expression, telomerase activity and change in the TERT gene promoter CpG island methylation status of human adipose-derived mesenchymal stem cells

“…The medium was replaced with a fresh medium every 3–4 days during cultivation. When the cells became confluent, they were trypsinized with 0.25% trypsin (Gibco, UK) and 1 mM EDTA (Invitrogen, UK), and then sub-cultured; hADSCs at Passage 3–6 were used in this study [ 26 , 27 ].…”

Natural Killer Cells from the Subcutaneous Adipose Tissue Underexpress the NKp30 and NKp44 in Obese Persons and Are Less Active against Major Histocompatibility Complex Class I Non-Expressing Neoplastic Cells

“…Also, the peripheral blood (PB) was obtained from these volunteers for the comparison between peripheral blood-derived NK cells (PBNKs) and adipose tissue-derived NK cells (ADNKs). SAT (20 ml for obese persons and only 10 ml for non-obese persons) was subjected to collagenase digestion, as described previously ( 25 ) to obtain the stromal vascular fraction (SVF). Mononuclear cells were isolated from SVF and also PB by Ficoll Paque centrifugation.…”

Comparison of intraoperative procedures for isolation of clinical grade stromal vascular fraction for regenerative purposes: a systematic review

“…Three studies were excluded based on the use of culture methods to isolate ASCs, because culture methods are incompatible with intraoperative applications (Busser et al, 2014;Priya et al, 2014;Wu et al, 2012). Four studies used only centrifugation or RBC lysis buffer as isolation protocol and were thereby excluded (Amirkhani et al, 2016;Baptista et al, 2009;Markarian et al, 2014;Raposio et al, 2014). Three studies used the blood saline fraction of lipoaspirate and were thereby excluded (Cicione et al, 2016;Francis et al, 2010;Shah et al, 2013).…”