Abstract
To investigate the differentiation potential of purified CD90+ cells sorted from adipose-derived stem cells (ADSCs), CD90+ cells were sorted from rabbit ADSCs using flow cytometry. Then, cell expansion of CD90+ cells and unsorted ADSCs was observed using an inverted microscope. Furthermore, cell surface markers including CD40, CD105, and CD90 on CD90+ cells and unsorted ADSCs were quantified using flow cytometry. Additionally, multi-lineage differentiation ability between CD90+ cells and unsorted ADSCs was compared, and expression of adipocyte-related genes PPAR-r and CEBPA as well as stem cell-related gene SOX2 in CD90+ cells and unsorted ADSCs was determined using real-time quantitative PCR. We found that CD90+ cells had a stronger cell proliferation ability than unsorted ADSCs. CD90+ cells showed a stronger ability of osteoblast and chondrocyte differentiation than unsorted ADSCs and CD90− cells, whereas the adipose differentiation ability of CD90+ cells was similar to that of ADSCs and CD90− cells. CD14, CD105, and CD90 on CD90+ cells were expressed more highly than those on ADSCs. Additionally, the mRNA expression level of SOX2 in CD90+ cells was significantly higher than that in ADSCs, whereas the expression of PPAR-r and CEBPA was markedly lower than that in ADSCs. These results suggested that the purified CD90+ cells sorted from ADSCs exhibit a stronger differentiation potential than the unsorted ADSCs.
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Aslan H, Zilberman Y, Kandel L, Liebergall M, Oskouian RJ, Gazit D, Gazit Z (2006) Osteogenic differentiation of noncultured immunoisolated bone marrow-derived CD105+ cells. Stem Cells 24:1728–1737
Braun J, Hack A, Weis-Klemm M, Conrad S, Treml S, Kohler K, Walliser U, Skutella T, Aicher WK (2010) Evaluation of the osteogenic and chondrogenic differentiation capacities of equine adipose tissue-derived mesenchymal stem cells. Am J Vet Res 71:1228–1236
Chen XD, Qian HY, Neff L, Satomura K, Horowitz MC (1999) Thy-1 antigen expression by cells in the osteoblast lineage. J Bone Miner Res 14:362–375
Chung MT, Liu C, Hyun JS, Lo DD, Montoro DT, Hasegawa M, Li S, Sorkin M, Rennert R, Keeney M (2013) CD90 (Thy-1)-positive selection enhances osteogenic capacity of human adipose-derived stromal cells. Tissue Eng Part A 19:989–997
Davies OG, Cooper PR, Shelton RM, Smith AJ, Scheven BA (2015) Isolation of adipose and bone marrow mesenchymal stem cells using CD29 and CD90 modifies their capacity for osteogenic and adipogenic differentiation. J Tissue Eng 6:2041731415592356
Dennis JE, Esterly K, Awadallah A, Parrish CR, Poynter GM, Goltry KL (2007) Clinical-scale expansion of a mixed population of bone marrow-derived stem and progenitor cells for potential use in bone tissue regeneration. Stem Cells 25:2575–2582
Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement Cytotherapy 8:315–317
García-Contreras M, Vera-Donoso CD, Hernandez-Andreu JM, García-Verdugo JM, Oltra E (2014) Therapeutic potential of human adipose-derived stem cells (ADSCs) from cancer patients: a pilot study. PLoS One 9:e113288
Gimble J, Rad MR, Yao S (2013) Adipose tissue-derived stem cells and their regeneration potential. Stem Cells Craniofac Dev Regen 241–258
Golipoor Z, Ragerdi Kashani I, Akbari M, Hassanzadeh G, Malek F, Mahmoudi R (2010) Differentiation of adipose-derived stem cells into schwann cell phenotype in comparison with bone marrow stem cells. Iranian Journal of Basic Medical Sciences 13:76–84
Hosoya A, Hiraga T, Ninomiya T, Yukita A, Yoshiba K, Yoshiba N, Takahashi M, Ito S, Nakamura H (2012) Thy-1-positive cells in the subodontoblastic layer possess high potential to differentiate into hard tissue-forming cells. Histochem Cell Biol 137:733–742
Ishimura D, Yamamoto N, Tajima K, Ohno A, Yamamoto Y, Washimi O, Yamada H (2008) Differentiation of adipose-derived stromal vascular fraction culture cells into chondrocytes using the method of cell sorting with a mesenchymal stem cell marker. Tohoku J Exp Med 216:149–156
Jiang T, Liu W, Lv X, Sun H, Zhang L, Liu Y, Zhang WJ, Cao Y, Zhou G (2010) Potent in vitro chondrogenesis of CD105 enriched human adipose-derived stem cells. Biomaterials 31:3564–3571
Krawetz RJ, Wu YE, Martin L, Rattner JB, Matyas JR, Hart DA (2012) Synovial fluid progenitors expressing CD90+ from normal but not osteoarthritic joints undergo chondrogenic differentiation without micro-mass culture. PLoS One 7:e43616
Liu K, Lin B, Zhao M, Yang X, Chen M, Gao A, Liu F, Que J, Lan X (2013) The multiple roles for Sox2 in stem cell maintenance and tumorigenesis. Cell Signal 25:1264–1271
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408
Miranville A, Heeschen C, Sengenes C, Curat C, Busse R, Bouloumie A (2004) Improvement of postnatal neovascularization by human adipose tissue-derived stem cells. Circulation 110:349–355
Mitterberger MC, Lechner S, Mattesich M, Kaiser A, Probst D, Wenger N, Pierer G, Zwerschke W (2012) DLK1 (PREF1) is a negative regulator of adipogenesis in CD105+/CD90+/CD34+/CD31−/FABP4− adipose-derived stromal cells from subcutaneous abdominal fat pats of adult women. Stem Cell Res 9:35–48
Niemelä S, Miettinen S, Sarkanen J, Ashammakhi N (2008) Adipose tissue and adipocyte differentiation: molecular and cellular aspects and tissue engineering applications. Topics in Tissue Engineering 4:26
Ramirez-Zacarias J, Castro-Munozledo F, Kuri-Harcuch W (1992) Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 97:493–497
Reich CM, Raabe O, Wenisch S, Bridger PS, Kramer M, Arnhold S (2012) Isolation, culture and chondrogenic differentiation of canine adipose tissue-and bone marrow-derived mesenchymal stem cells—a comparative study. Vet Res Commun 36:139–148
Shin S, Kim Y, Jeong S, Hong S, Kim I, Lee W, Choi S (2013) The therapeutic effect of human adult stem cells derived from adipose tissue in endotoxemic rat model. International journal of medical sciences 10:8
Yamamoto M, Nakata H, Hao J, Chou J, Kasugai S, Kuroda S (2014) Osteogenic potential of mouse adipose-derived stem cells sorted for CD90 and CD105 in vitro. Stem Cells Int 2014:1–17
Yanez R, Lamana ML, García-Castro J, Colmenero I, Ramírez M, Bueren JA (2006) Adipose tissue-derived mesenchymal stem cells have in vivo immunosuppressive properties applicable for the control of the graft-versus-host disease. Stem Cells 24:2582–2591
Acknowledgments
The authors are grateful to the Core Facilities of Zhejiang University School of Medicine for technical assistance. This work was supported by the Project of Science and Technology Department of Zhejiang Province (no. 2014C37067, no. 2015C37008), the National Natural Science Foundation of China (no. 81501943), the Fundamental Research Funds for the Central Universities (no. 2016QNA7021), and the Important Subject Fund of Science Technology Department of Zhejiang province (no. 2013C03048-1).
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Editor: Tetsuji Okamoto
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Song, X., Hong, C., Zheng, Q. et al. Differentiation potential of rabbit CD90-positive cells sorted from adipose-derived stem cells in vitro. In Vitro Cell.Dev.Biol.-Animal 53, 77–82 (2017). https://doi.org/10.1007/s11626-016-0081-6
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DOI: https://doi.org/10.1007/s11626-016-0081-6