Wet milling of large quantities of human excision adipose tissue for the isolation of stromal vascular fraction cells
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The isolation of stromal vascular fraction (SVF) cells from excised human adipose tissue, for clinical or research purposes, implies the tedious and time consuming process of manual mincing prior to enzymatic digestion. Since no efficient alternative technique to this current standard procedure has been proposed so far, the aim of this study was to test a milling procedure, using two simple, inexpensive and commercially available manual meat grinders, to process large amounts of adipose tissue. The procedure was assessed on adipose tissue resections from seven human donors and compared to manual mincing with scalpels. The processed adipose tissues were digested and the resulting SVF cells compared in terms of number, clonogenicity and differentiation capacity. After 10 min of processing, either device tested yielded on average sixfold more processed material for subsequent cell isolation than manual mincing. The isolation yield of SVF cells (isolated cells per ml of adipose tissue), their viability, phenotype, clonogenicity and osteogenic/adipogenic differentiation capacity, tested by production of mineralized matrix and lipid vacuoles, respectively, were comparable. This new method is practical and inexpensive and represents an efficient alternative to the current standard for large scale adipose tissue resection processing. A device based on the milling principle could be embedded within a streamlined system for isolation and clinical use of SVF cells from adipose tissue excision.
KeywordsAdipose tissue derived stem cells Mesenchymal stem cells Stromal vascular fraction Mechanical device Isolation Human adipose tissue
This work was supported by the Swiss National Science Foundation (SNF Grant No. 310030-138519, to A.S. and I.M.).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was approved by the local ethical committee (Ethikkommission beider Basel [EKBB], Ref. 78/07 extended in 2009). All seven donors gave written informed consent.
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- CDC Guideline for disinfection and Sterilization in Healthcare Facilities (2008). https://www.cdc.gov/infectioncontrol/pdf/guidelines/disinfection-guidelines.pdf. Accessed 17 June 2017
- Fraser JK, Zhu M, Wulur I, Alfonso Z (2008) Adipose-derived stem cells. Methods Mol Biol 449:59–67Google Scholar
- Kølle SF, Fischer-Nielsen A, Mathiasen AB, Elberg JJ, Oliveri RS, Glovinski PV, Kastrup J, Kirchhoff M, Rasmussen BS, Talman ML, Thomsen C, Dickmeiss E, Drzewiecki KT (2013) Enrichment of autologous fat grafts with ex vivo expanded adipose tissue-derived stem cells for graft survival: a randomised placebo-controlled trial. Lancet 382:1113–1120CrossRefGoogle Scholar
- Mehrkens A, Saxer F, Güven S, Hoffmann W, Müller AM, Jakob M, Weber FE, Martin I, Scherberich A (2012) Intraoperative engineering of osteogenic grafts combining freshly harvested, human adipose-derived cells and physiological doses of bone morphogenetic protein-2. Eur Cell Mater 28:308–319CrossRefGoogle Scholar
- Müller AM, Mehrkens A, Schäfer DJ, Jaquiery C, Güven S, Lehmicke M, Martinetti R, Farhadi I, Jakob M, Scherberich A, Martin I (2010) Towards an intraoperative engineering of osteogenic and vasculogenic grafts from the stromal vascular fraction of human adipose tissue. Eur Cell Mater 3:127–135CrossRefGoogle Scholar
- Salem HK, Thiemermann C (2010) Mesenchymal stromal cells: current understanding and clinical status. Stem Cells 31:585–596Google Scholar
- Saxer F, Scherberich A, Todorov A, Studer P, Miot S, Schreiner S, Güven S, Tchang LA, Haug M, Heberer M, Schaefer DJ, Rikli D, Martin I, Jakob M (2016) Implantation of stromal vascular fraction progenitors at bone fracture sites: from a rat model to a first-in-man study. Stem Cells 34:2956–2966. https://doi.org/10.1002/stem.2478 CrossRefGoogle Scholar
- Volat F, Bouloumié A (2013) Steroid hormones and the stroma-vascular cells of the adipose tissue. Horm Mol Biol Clin Investig 15:5–10Google Scholar