Skip to main content
Top
Published in: European Journal of Medical Research 1/2017

Open Access 01-12-2017 | Research

Adipose-derived mesenchymal stem cells from liposuction and resected fat are feasible sources for regenerative medicine

Authors: Sandra Schneider, Marina Unger, Martijn van Griensven, Elizabeth R. Balmayor

Published in: European Journal of Medical Research | Issue 1/2017

Login to get access

Abstract

Background

The use of mesenchymal stem cells (MSCs) in research and in regenerative medicine has progressed. Bone marrow as a source has drawbacks because of subsequent morbidities. An easily accessible and valuable source is adipose tissue. This type of tissue contains a high number of MSCs, and obtaining higher quantities of tissue is more feasible. Fat tissue can be harvested using different methods such as liposuction and resection. First, a detailed isolation protocol with complete characterization is described. This also includes highlighting problems and pitfalls. Furthermore, some comparisons of these different harvesting methods exist. However, the later characterization of the cells is conducted poorly in most cases.

Methods

We performed an in-depth characterization over five passages including an investigation of the effect of freezing and thawing. Characterization was performed using flow cytometry with CD markers, metabolic activity with Alamar Blue, growth potential in between passages, and cytoskeleton staining.

Results

Our results show that the cells isolated with distinct isolation methods (solid versus liposuction “liquid”) have the same MSC potential. However, the percentage of cells positive for the markers CD73, CD90, and CD105 is initially quite low. The cells isolated from the liquid fat tissue grow faster at higher passages, and significantly more cells display MSC markers.

Conclusion

In summary, we show a simple and efficient method to isolate adipose-derived mesenchymal stem cells from different preparations. Liposuctions and resection can be used, whereas liposuction has more growth potential at higher passages.
Literature
1.
go back to reference Baer PC, Geiger H. Adipose-derived mesenchymal stromal/stem cells: tissue localization, characterization, and heterogeneity. Stem Cells Int. 2012;2012:812693.CrossRefPubMedPubMedCentral Baer PC, Geiger H. Adipose-derived mesenchymal stromal/stem cells: tissue localization, characterization, and heterogeneity. Stem Cells Int. 2012;2012:812693.CrossRefPubMedPubMedCentral
2.
go back to reference Hynes K, Menicanin D, Mrozik K, Gronthos S, Bartold PM. Generation of functional mesenchymal stem cells from different induced pluripotent stem cell lines. Stem Cells Dev. 2014;23:1084–96.CrossRefPubMed Hynes K, Menicanin D, Mrozik K, Gronthos S, Bartold PM. Generation of functional mesenchymal stem cells from different induced pluripotent stem cell lines. Stem Cells Dev. 2014;23:1084–96.CrossRefPubMed
3.
go back to reference Otte A, Bucan V, Reimers K, Hass R. Mesenchymal stem cells maintain long-term in vitro stemness during explant culture. Tissue Eng Part C Methods. 2013;19:937–48.CrossRefPubMed Otte A, Bucan V, Reimers K, Hass R. Mesenchymal stem cells maintain long-term in vitro stemness during explant culture. Tissue Eng Part C Methods. 2013;19:937–48.CrossRefPubMed
4.
go back to reference Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7:211–28.CrossRefPubMed Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7:211–28.CrossRefPubMed
5.
go back to reference Faustini M, Bucco M, Chlapanidas T, Lucconi G, Marazzi M, Tosca MC, Gaetani P, Klinger M, Villani S, Ferretti VV, et al. Nonexpanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues. Tissue Eng Part C Methods. 2010;16:1515–21.CrossRefPubMed Faustini M, Bucco M, Chlapanidas T, Lucconi G, Marazzi M, Tosca MC, Gaetani P, Klinger M, Villani S, Ferretti VV, et al. Nonexpanded mesenchymal stem cells for regenerative medicine: yield in stromal vascular fraction from adipose tissues. Tissue Eng Part C Methods. 2010;16:1515–21.CrossRefPubMed
6.
go back to reference Eto H, Suga H, Matsumoto D, Inoue K, Aoi N, Kato H, Araki J, Yoshimura K. Characterization of structure and cellular components of aspirated and excised adipose tissue. Plast Reconstr Surg. 2009;124:1087–97.CrossRefPubMed Eto H, Suga H, Matsumoto D, Inoue K, Aoi N, Kato H, Araki J, Yoshimura K. Characterization of structure and cellular components of aspirated and excised adipose tissue. Plast Reconstr Surg. 2009;124:1087–97.CrossRefPubMed
7.
go back to reference Busser H, Najar M, Raicevic G, Pieters K, Velez Pombo R, Philippart P, Meuleman N, Bron D, Lagneaux L. Isolation and characterization of human mesenchymal stromal cell subpopulations: comparison of bone marrow and adipose tissue. Stem Cells Dev. 2015;24:2142–57.CrossRefPubMed Busser H, Najar M, Raicevic G, Pieters K, Velez Pombo R, Philippart P, Meuleman N, Bron D, Lagneaux L. Isolation and characterization of human mesenchymal stromal cell subpopulations: comparison of bone marrow and adipose tissue. Stem Cells Dev. 2015;24:2142–57.CrossRefPubMed
8.
go back to reference Ardeshirylajimi A, Rafeie F, Zandi-Karimi A, Jaffarabadi GA, Mohammadi-Sangcheshmeh A, Samiei R, Toghdory A, Seyedjafari E, Hashemi SM, Cinar MU, Gastal EL. Fat harvesting site is an important determinant of proliferation and pluripotency of adipose-derived stem cells. Biologicals. 2016;44:12–8.CrossRefPubMed Ardeshirylajimi A, Rafeie F, Zandi-Karimi A, Jaffarabadi GA, Mohammadi-Sangcheshmeh A, Samiei R, Toghdory A, Seyedjafari E, Hashemi SM, Cinar MU, Gastal EL. Fat harvesting site is an important determinant of proliferation and pluripotency of adipose-derived stem cells. Biologicals. 2016;44:12–8.CrossRefPubMed
9.
go back to reference Jung S, Kleineidam B, Kleinheinz J. Regenerative potential of human adipose-derived stromal cells of various origins. J Craniomaxillofac Surg. 2015;43:2144–51.CrossRefPubMed Jung S, Kleineidam B, Kleinheinz J. Regenerative potential of human adipose-derived stromal cells of various origins. J Craniomaxillofac Surg. 2015;43:2144–51.CrossRefPubMed
10.
go back to reference Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, Zandiehdoulabi B, Schouten TE, Kuik DJ, Ritt MJ, van Milligen FJ. Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res. 2008;332:415–26.CrossRefPubMedPubMedCentral Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, Zandiehdoulabi B, Schouten TE, Kuik DJ, Ritt MJ, van Milligen FJ. Effect of tissue-harvesting site on yield of stem cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res. 2008;332:415–26.CrossRefPubMedPubMedCentral
11.
12.
go back to reference Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7.CrossRefPubMed Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7.CrossRefPubMed
13.
go back to reference Baptista LS, do Amaral RJ, Carias RB, Aniceto M, Claudio-da-Silva C, Borojevic R. An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples. Cytotherapy. 2009;11:706–15.CrossRefPubMed Baptista LS, do Amaral RJ, Carias RB, Aniceto M, Claudio-da-Silva C, Borojevic R. An alternative method for the isolation of mesenchymal stromal cells derived from lipoaspirate samples. Cytotherapy. 2009;11:706–15.CrossRefPubMed
14.
go back to reference Bony C, Cren M, Domergue S, Toupet K, Jorgensen C, Noel D. Adipose mesenchymal stem cells isolated after manual or water-jet-assisted liposuction display similar properties. Front Immunol. 2015;6:655.PubMed Bony C, Cren M, Domergue S, Toupet K, Jorgensen C, Noel D. Adipose mesenchymal stem cells isolated after manual or water-jet-assisted liposuction display similar properties. Front Immunol. 2015;6:655.PubMed
15.
go back to reference Chaput B, Bertheuil N, Escubes M, Grolleau JL, Garrido I, Laloze J, Espagnolle N, Casteilla L, Sensebe L, Varin A. Mechanically isolated stromal vascular fraction provides a valid and useful collagenase-free alternative technique: a comparative study. Plast Reconstr Surg. 2016;138:807–19.CrossRefPubMed Chaput B, Bertheuil N, Escubes M, Grolleau JL, Garrido I, Laloze J, Espagnolle N, Casteilla L, Sensebe L, Varin A. Mechanically isolated stromal vascular fraction provides a valid and useful collagenase-free alternative technique: a comparative study. Plast Reconstr Surg. 2016;138:807–19.CrossRefPubMed
16.
go back to reference Domenis R, Lazzaro L, Calabrese S, Mangoni D, Gallelli A, Bourkoula E, Manini I, Bergamin N, Toffoletto B, Beltrami CA, et al. Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques. Stem Cell Res Ther. 2015;6:2.CrossRefPubMedPubMedCentral Domenis R, Lazzaro L, Calabrese S, Mangoni D, Gallelli A, Bourkoula E, Manini I, Bergamin N, Toffoletto B, Beltrami CA, et al. Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques. Stem Cell Res Ther. 2015;6:2.CrossRefPubMedPubMedCentral
17.
go back to reference Gittel C, Brehm W, Burk J, Juelke H, Staszyk C, Ribitsch I. Isolation of equine multipotent mesenchymal stromal cells by enzymatic tissue digestion or explant technique: comparison of cellular properties. BMC Vet Res. 2013;9:221.CrossRefPubMedPubMedCentral Gittel C, Brehm W, Burk J, Juelke H, Staszyk C, Ribitsch I. Isolation of equine multipotent mesenchymal stromal cells by enzymatic tissue digestion or explant technique: comparison of cellular properties. BMC Vet Res. 2013;9:221.CrossRefPubMedPubMedCentral
18.
go back to reference Guven S, Karagianni M, Schwalbe M, Schreiner S, Farhadi J, Bula S, Bieback K, Martin I, Scherberich A. Validation of an automated procedure to isolate human adipose tissue-derived cells by using the Sepax(R) technology. Tissue Eng Part C Methods. 2012;18:575–82.CrossRefPubMedPubMedCentral Guven S, Karagianni M, Schwalbe M, Schreiner S, Farhadi J, Bula S, Bieback K, Martin I, Scherberich A. Validation of an automated procedure to isolate human adipose tissue-derived cells by using the Sepax(R) technology. Tissue Eng Part C Methods. 2012;18:575–82.CrossRefPubMedPubMedCentral
19.
go back to reference Najar M, Rodrigues RM, Buyl K, Branson S, Vanhaecke T, Lagneaux L, Rogiers V, De Kock J. Proliferative and phenotypical characteristics of human adipose tissue-derived stem cells: comparison of Ficoll gradient centrifugation and red blood cell lysis buffer treatment purification methods. Cytotherapy. 2014;16:1220–8.CrossRefPubMed Najar M, Rodrigues RM, Buyl K, Branson S, Vanhaecke T, Lagneaux L, Rogiers V, De Kock J. Proliferative and phenotypical characteristics of human adipose tissue-derived stem cells: comparison of Ficoll gradient centrifugation and red blood cell lysis buffer treatment purification methods. Cytotherapy. 2014;16:1220–8.CrossRefPubMed
20.
go back to reference Ong WK, Tan CS, Chan KL, Goesantoso GG, Chan XH, Chan E, Yin J, Yeo CR, Khoo CM, So JB, et al. Identification of specific cell-surface markers of adipose-derived stem cells from subcutaneous and visceral fat depots. Stem Cell Rep. 2014;2:171–9.CrossRef Ong WK, Tan CS, Chan KL, Goesantoso GG, Chan XH, Chan E, Yin J, Yeo CR, Khoo CM, So JB, et al. Identification of specific cell-surface markers of adipose-derived stem cells from subcutaneous and visceral fat depots. Stem Cell Rep. 2014;2:171–9.CrossRef
21.
go back to reference Priya N, Sarcar S, Majumdar AS, SundarRaj S. Explant culture: a simple, reproducible, efficient and economic technique for isolation of mesenchymal stromal cells from human adipose tissue and lipoaspirate. J Tissue Eng Regen Med. 2014;8:706–16.CrossRefPubMed Priya N, Sarcar S, Majumdar AS, SundarRaj S. Explant culture: a simple, reproducible, efficient and economic technique for isolation of mesenchymal stromal cells from human adipose tissue and lipoaspirate. J Tissue Eng Regen Med. 2014;8:706–16.CrossRefPubMed
22.
go back to reference Zhu M, Heydarkhan-Hagvall S, Hedrick M, Benhaim P, Zuk P. Manual isolation of adipose-derived stem cells from human lipoaspirates. J Vis Exp. 2013;79:e50585. Zhu M, Heydarkhan-Hagvall S, Hedrick M, Benhaim P, Zuk P. Manual isolation of adipose-derived stem cells from human lipoaspirates. J Vis Exp. 2013;79:e50585.
23.
go back to reference Estes BT, Diekman BO, Gimble JM, Guilak F. Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype. Nat Protoc. 2010;5:1294–311.CrossRefPubMedPubMedCentral Estes BT, Diekman BO, Gimble JM, Guilak F. Isolation of adipose-derived stem cells and their induction to a chondrogenic phenotype. Nat Protoc. 2010;5:1294–311.CrossRefPubMedPubMedCentral
24.
go back to reference Minonzio G, Corazza M, Mariotta L, Gola M, Zanzi M, Gandolfi E, De Fazio D, Soldati G. Frozen adipose-derived mesenchymal stem cells maintain high capability to grow and differentiate. Cryobiology. 2014;69:211–6.CrossRefPubMed Minonzio G, Corazza M, Mariotta L, Gola M, Zanzi M, Gandolfi E, De Fazio D, Soldati G. Frozen adipose-derived mesenchymal stem cells maintain high capability to grow and differentiate. Cryobiology. 2014;69:211–6.CrossRefPubMed
25.
go back to reference Devitt SM, Carter CM, Dierov R, Weiss S, Gersch RP, Percec I. Successful isolation of viable adipose-derived stem cells from human adipose tissue subject to long-term cryopreservation: positive implications for adult stem cell-based therapeutics in patients of advanced age. Stem Cells Int. 2015;2015:146421.CrossRefPubMedPubMedCentral Devitt SM, Carter CM, Dierov R, Weiss S, Gersch RP, Percec I. Successful isolation of viable adipose-derived stem cells from human adipose tissue subject to long-term cryopreservation: positive implications for adult stem cell-based therapeutics in patients of advanced age. Stem Cells Int. 2015;2015:146421.CrossRefPubMedPubMedCentral
Metadata
Title
Adipose-derived mesenchymal stem cells from liposuction and resected fat are feasible sources for regenerative medicine
Authors
Sandra Schneider
Marina Unger
Martijn van Griensven
Elizabeth R. Balmayor
Publication date
01-12-2017
Publisher
BioMed Central
Published in
European Journal of Medical Research / Issue 1/2017
Electronic ISSN: 2047-783X
DOI
https://doi.org/10.1186/s40001-017-0258-9

Other articles of this Issue 1/2017

European Journal of Medical Research 1/2017 Go to the issue