Comparative analysis of human mesenchymal stem cells from bone marrow and adipose tissue under xeno-free conditions for cell therapy

Chun-yu Li, Xiao-yun Wu, Jia-bei Tong, Xin-xin Yang, Jing-li Zhao, Quan-fu Zheng, Guo-bin Zhao, Zhi-jie Ma, Chun-yu Li, Xiao-yun Wu, Jia-bei Tong, Xin-xin Yang, Jing-li Zhao, Quan-fu Zheng, Guo-bin Zhao, Zhi-jie Ma

Abstract

Introduction: Mesenchymal stem cells (MSCs) are promising candidates for cell-based therapies. Human platelet lysate represents an efficient alternative to fetal bovine serum for clinical-scale expansion of MSCs. Different media used in culture processes should maintain the biological characteristics of MSCs during multiple passages. However, bone marrow-derived MSCs and adipose tissue-derived MSCs have not yet been directly compared with each other under human platelet lysate conditions. This study aims to conduct a direct head-to-head comparison of the biological characteristics of the two types of MSCs under human platelet lysate-supplemented culture conditions for their ability to be used in regenerative medicine applications.

Methods: The bone marrow- and adipose tissue-derived MSCs were cultured under human platelet lysate conditions and their biological characteristics evaluated for cell therapy (morphology, immunophenotype, colony-forming unit-fibroblast efficiency, proliferation capacity, potential for mesodermal differentiation, secreted proteins, and immunomodulatory effects).

Results: Under human platelet lysate-supplemented culture conditions, bone marrow- and adipose tissue-derived MSCs exhibited similar fibroblast-like morphology and expression patterns of surface markers. Adipose tissue-derived MSCs had greater proliferative potential than bone marrow-derived MSCs, while no significantly difference in colony efficiency were observed between the two types of cells. However, bone marrow-derived MSCs possessed higher capacity toward osteogenic and chondrogenic differentiation compared with adipose tissue-derived MSCs, while similar adipogenic differentiation potential wase observed between the two types of cells. There were some differences between bone marrow- and adipose tissue-derived MSCs for several secreted proteins, such as cytokine (interferon-γ), growth factors (basic fibroblast growth factor, hepatocyte growth factor, and insulin-like growth factor-1), and chemokine (stem cell-derived factor-1). Adipose tissue-derived MSCs had more potent immunomodulatory effects than bone marrow-derived MSCs.

Conclusions: Adipose tissue-derived MSCs have biological advantages in the proliferative capacity, secreted proteins (basic fibroblast growth factor, interferon-γ, and insulin-like growth factor-1), and immunomodulatory effects, but bone marrow-derived MSCs have advantages in osteogenic and chondrogenic differentiation potential and secreted proteins (stem cell-derived factor-1 and hepatocyte growth factor); these biological advantages should be considered systematically when choosing the MSC source for specific clinical application.

Figures

Figure 1
Figure 1
Morphology of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells cultivated in human platelet lysate-supplemented medium. BMMSCs and ATMSCs were tiny, slender, and bright. Scale bar = 100 μm.
Figure 2
Figure 2
Proliferation capacity of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells in human platelet lysate-supplemented medium. (A) Cumulative population doublings of BMMSCs and ATMSCs. (B) Colony-forming unit-fibroblast (CFU-F) number of BMMSCs and ATMSCs cultivated in human platelet lysate-supplemented medium at densities of 1,000 per well. All values are shown as means ± standard deviation; n = 5. *P < 0.05, **P < 0.01. n.s., not significant.
Figure 3
Figure 3
Immunophenotype of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells in human platelet lysate-supplemented medium. Flow cytometry analysis showed that BMMSCs and ATMSCs consistently stained positive for CD13, CD29, CD90, CD105, and HLA-ABC and did not react with CD14, CD19, CD34, CD45, and HLA-DR (n = 5; one representative example shown).
Figure 4
Figure 4
Osteogenic differentiation potential of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells in human platelet lysate-supplemented medium. (A) Alizarin Red staining of BMMSCs and ATMSCs. Magnification × 100. (B) Quantitative assessment of mRNA expression. Values are shown as mean ± standard deviation; n = 5. *P < 0.05.
Figure 5
Figure 5
Chondrogenic differentiation potential of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells in human platelet lysate-supplemented medium. (A) Alcian Blue staining of BMMSCs and ATMSCs. Magnification × 100. (B) Quantitative assessment of mRNA expression. Values are shown as mean ± standard deviation; n = 5. **P < 0.01.
Figure 6
Figure 6
Adipogenic differentiation potential of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells in human platelet lysate-supplemented medium. (A) Oil Red O staining of BMMSCs and ATMSCs. Magnification × 200. (B) Quantitative assessment of mRNA expression. Values are shown as mean ± standard deviation; n = 5. n.s., not significant.
Figure 7
Figure 7
Quantification of cytokines, growth factors, and chemokines (pg/105 cells) of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells. Values are shown as mean ± standard deviation; n = 5. *P < 0.05, **P < 0.01. bFGF, basic fibroblast growth factor; HGF, hepatocyte growth factor; IFN, interferon; IGF, insulin-like growth factor; IL, interleukin; n.s., not significant; SDF-1, stem cell-derived factor-1; TNF, tumor necrosis factor; VEGF, vascular endothelial growth factor.
Figure 8
Figure 8
Quantification of immunomodulatory factor (pg/mL) of bone marrow- (BMMSCs) and adipose tissue (ATMSCs)-derived mesenchymal stem cells. Values are shown as mean ± standard deviation; n = 5. *P < 0.01. MSC, mesenchymal stem cell; n.s., not significant; PGE2, prostaglandin E2; TGF, transforming growth factor.

References

    1. Zou JP, Huang S, Peng Y, Liu HW, Cheng B, Fu XB, et al. Mesenchymal stem cells/multipotent mesenchymal stromal cells (MSCs): potential role in healing cutaneous chronic wounds. Int J Low Extrem Wounds. 2012;11:244–53.
    1. da Silva ML, Chagastelles PC, Nardi NB. Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J Cell Sci. 2006;119:2204–13. doi: 10.1242/jcs.02932.
    1. Hoogduijn MJ, Dor FJ. Mesenchymal stem cells: are we ready for clinical application in transplantation and tissue regeneration? Front Immunol. 2013;4:144. doi: 10.3389/fimmu.2013.00144.
    1. Puissant B, Barreau C, Bourin P, Clavel C, Corre J, Bousquet C, et al. Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells. Br J Haematol. 2005;129:118–29. doi: 10.1111/j.1365-2141.2005.05409.x.
    1. Chung CS, Fujita N, Kawahara N, Yui S, Nam E, Nishimura R. A comparison of neurosphere differentiation potential of canine bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells. J Vet Med Sci. 2013;75:879–86. doi: 10.1292/jvms.12-0470.
    1. Cooper GM, Durham EL, Cray JJ, Jr, Bykowski MR, DeCesare GE, Smalley MA, et al. Direct comparison of progenitor cells derived from adipose, muscle, and bone marrow from wild-type or craniosynostotic rabbits. Plast Reconstr Surg. 2011;127:88–97. doi: 10.1097/PRS.0b013e3181fad311.
    1. Danisovic L, Varga I, Polak S, Ulicna M, Hlavackova L, Bohmer D, et al. Comparison of in vitro chondrogenic potential of human mesenchymal stem cells derived from bone marrow and adipose tissue. Gen Physiol Biophys. 2009;28:56–62. doi: 10.4149/gpb_2009_01_56.
    1. De Ugarte DA, Morizono K, Elbarbary A, Alfonso Z, Zuk PA, Zhu M, et al. Comparison of multi-lineage cells from human adipose tissue and bone marrow. Cells Tissues Organs. 2003;174:101–9. doi: 10.1159/000071150.
    1. Vishnubalaji R, Al-Nbaheen M, Kadalmani B, Aldahmash A, Ramesh T. Comparative investigation of the differentiation capability of bone-marrow- and adipose-derived mesenchymal stem cells by qualitative and quantitative analysis. Cell Tissue Res. 2012;347:419–27. doi: 10.1007/s00441-011-1306-3.
    1. Elman JS, Li M, Wang F, Gimble JM, Parekkadan B. A comparison of adipose and bone marrow-derived mesenchymal stromal cell secreted factors in the treatment of systemic inflammation. J Inflamm. 2014;11:1. doi: 10.1186/1476-9255-11-1.
    1. Ertas G, Ural E, Ural D, Aksoy A, Kozdag G, Gacar G, et al. Comparative analysis of apoptotic resistance of mesenchymal stem cells isolated from human bone marrow and adipose tissue. ScientificWorldJournal. 2012;2012:105698. doi: 10.1100/2012/105698.
    1. Rasmussen JG, Frobert O, Holst-Hansen C, Kastrup J, Baandrup U, Zachar V, et al. Comparison of human adipose-derived stem cells and bone marrow-derived stem cells in a myocardial infarction model. Cell Transplant. 2014;23:195–206. doi: 10.3727/096368912X659871.
    1. Huang JI, Kazmi N, Durbhakula MM, Hering TM, Yoo JU, Johnstone B. Chondrogenic potential of progenitor cells derived from human bone marrow and adipose tissue: a patient-matched comparison. J Orthop Res. 2005;23:1383–9. doi: 10.1016/j.orthres.2005.03.008.1100230621.
    1. Hsiao ST, Asgari A, Lokmic Z, Sinclair R, Dusting GJ, Lim SY, et al. Comparative analysis of paracrine factor expression in human adult mesenchymal stem cells derived from bone marrow, adipose, and dermal tissue. Stem Cells Dev. 2012;21:2189–203. doi: 10.1089/scd.2011.0674.
    1. Ahmadian Kia N, Bahrami AR, Ebrahimi M, Matin MM, Neshati Z, Almohaddesin MR, et al. Comparative analysis of chemokine receptor's expression in mesenchymal stem cells derived from human bone marrow and adipose tissue. J Mol Neurosci. 2011;44:178–85. doi: 10.1007/s12031-010-9446-6.
    1. Muller I, Kordowich S, Holzwarth C, Spano C, Isensee G, Staiber A, et al. Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM. Cytotherapy. 2006;8:437–44. doi: 10.1080/14653240600920782.
    1. van der Valk J, Brunner D, De Smet K, Fex Svenningsen A, Honegger P, Knudsen LE, et al. Optimization of chemically defined cell culture media–replacing fetal bovine serum in mammalian in vitro methods. Toxicol In Vitro. 2010;24:1053–63. doi: 10.1016/j.tiv.2010.03.016.
    1. Spees JL, Gregory CA, Singh H, Tucker HA, Peister A, Lynch PJ, et al. Internalized antigens must be removed to prepare hypoimmunogenic mesenchymal stem cells for cell and gene therapy. Mol Ther. 2004;9:747–56. doi: 10.1016/j.ymthe.2004.02.012.
    1. Lange C, Cakiroglu F, Spiess AN, Cappallo-Obermann H, Dierlamm J, Zander AR. Accelerated and safe expansion of human mesenchymal stromal cells in animal serum-free medium for transplantation and regenerative medicine. J Cell Physiol. 2007;213:18–26. doi: 10.1002/jcp.21081.
    1. Capelli C, Domenghini M, Borleri G, Bellavita P, Poma R, Carobbio A, et al. Human platelet lysate allows expansion and clinical grade production of mesenchymal stromal cells from small samples of bone marrow aspirates or marrow filter washouts. Bone Marrow Transplant. 2007;40:785–91. doi: 10.1038/sj.bmt.1705798.
    1. Centeno CJ, Schultz JR, Cheever M, Freeman M, Faulkner S, Robinson B, et al. Safety and complications reporting update on the re-implantation of culture-expanded mesenchymal stem cells using autologous platelet lysate technique. Curr Stem Cell Res Ther. 2011;6:368–78. doi: 10.2174/157488811797904371.
    1. Naaijkens BA, Niessen HW, Prins HJ, Krijnen PA, Kokhuis TJ, de Jong N, et al. Human platelet lysate as a fetal bovine serum substitute improves human adipose-derived stromal cell culture for future cardiac repair applications. Cell Tissue Res. 2012;348:119–30. doi: 10.1007/s00441-012-1360-5.
    1. Schallmoser K, Bartmann C, Rohde E, Reinisch A, Kashofer K, Stadelmeyer E, et al. Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells. Transfusion. 2007;47:1436–46. doi: 10.1111/j.1537-2995.2007.01220.x.
    1. Walenda G, Hemeda H, Schneider RK, Merkel R, Hoffmann B, Wagner W. Human platelet lysate gel provides a novel three dimensional-matrix for enhanced culture expansion of mesenchymal stromal cells. Tissue Eng Part C Methods. 2012;18:924–34. doi: 10.1089/ten.tec.2011.0541.
    1. Gottipamula S, Sharma A, Krishnamurthy S, Majumdar AS, Seetharam RN. Human platelet lysate is an alternative to fetal bovine serum for large-scale expansion of bone marrow-derived mesenchymal stromal cells. Biotechnol Lett. 2012;34:1367–74. doi: 10.1007/s10529-012-0893-8.
    1. Xia W, Li H, Wang Z, Xu R, Fu Y, Zhang X, et al. Human platelet lysate supports ex vivo expansion and enhances osteogenic differentiation of human bone marrow-derived mesenchymal stem cells. Cell Biol Int. 2011;35:639–43. doi: 10.1042/CBI20100361.
    1. Abdelrazik H, Spaggiari GM, Chiossone L, Moretta L. Mesenchymal stem cells expanded in human platelet lysate display a decreased inhibitory capacity on T- and NK-cell proliferation and function. Eur J Immunol. 2011;41:3281–90. doi: 10.1002/eji.201141542.
    1. Sensebe L, Bourin P, Tarte K. Good manufacturing practices production of mesenchymal stem/stromal cells. Hum Gene Ther. 2011;22:19–26. doi: 10.1089/hum.2010.197.
    1. Meirelles Lda S, Fontes AM, Covas DT, Caplan AI. Mechanisms involved in the therapeutic properties of mesenchymal stem cells. Cytokine Growth Factor Rev. 2009;20:419–27. doi: 10.1016/j.cytogfr.2009.10.002.
    1. Mok PL, Leong CF, Cheong SK. Cellular mechanisms of emerging applications of mesenchymal stem cells. Malays J Pathol. 2013;35:17–32.
    1. Ozaki K, Sato K, Oh I, Meguro A, Tatara R, Muroi K, et al. Mechanisms of immunomodulation by mesenchymal stem cells. Int J Hematol. 2007;86:5–7. doi: 10.1532/IJH97.07003.
    1. Schallmoser K, Rohde E, Reinisch A, Bartmann C, Thaler D, Drexler C, et al. Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum. Tissue Eng Part C Methods. 2008;14:185–96. doi: 10.1089/ten.tec.2008.0060.
    1. Parker AM, Shang H, Khurgel M, Katz AJ. Low serum and serum-free culture of multipotential human adipose stem cells. Cytotherapy. 2007;9:637–46. doi: 10.1080/14653240701508452.
    1. Yoshimura H, Muneta T, Nimura A, Yokoyama A, Koga H, Sekiya I. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res. 2007;327:449–62. doi: 10.1007/s00441-006-0308-z.
    1. Zimmermann JA, McDevitt TC. Pre-conditioning mesenchymal stromal cell spheroids for immunomodulatory paracrine factor secretion. Cytotherapy. 2014;16:331–45. doi: 10.1016/j.jcyt.2013.09.004.
    1. Abdelkhalek NK, Komiya A, Kato-Unoki Y, Somamoto T, Nakao M. Molecular evidence for the existence of two distinct IL-8 lineages of teleost CXC-chemokines. Fish Shellfish Immunol. 2009;27:763–7. doi: 10.1016/j.fsi.2009.08.004.
    1. Zhou Y, Guan X, Yu M, Wang X, Zhu W, Wang C, et al. Angiogenic/osteogenic response of BMMSCs on bone-derived scaffold: effect of hypoxia and role of PI3K/Akt-mediated VEGF-VEGFR pathway. Biotechnol J. 2014.
    1. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315–7. doi: 10.1080/14653240600855905.
    1. Mosna F, Sensebe L, Krampera M. Human bone marrow and adipose tissue mesenchymal stem cells: a user's guide. Stem Cells Dev. 2010;19:1449–70. doi: 10.1089/scd.2010.0140.
    1. Secunda R, Vennila R, Mohanashankar AM, Rajasundari M, Jeswanth S, Surendran R. Isolation, expansion and characterisation of mesenchymal stem cells from human bone marrow, adipose tissue, umbilical cord blood and matrix: a comparative study. Cytotechnology. 2014. [Epub ahead of print].
    1. Jin HJ, Bae YK, Kim M, Kwon SJ, Jeon HB, Choi SJ, et al. Comparative analysis of human mesenchymal stem cells from bone marrow, adipose tissue, and umbilical cord blood as sources of cell therapy. Int J Mol Sci. 2013;14:17986–8001. doi: 10.3390/ijms140917986.
    1. Peng L, Jia Z, Yin X, Zhang X, Liu Y, Chen P, et al. Comparative analysis of mesenchymal stem cells from bone marrow, cartilage, and adipose tissue. Stem Cells Dev. 2008;17:761–73. doi: 10.1089/scd.2007.0217.
    1. Nakanishi C, Nagaya N, Ohnishi S, Yamahara K, Takabatake S, Konno T, et al. Gene and protein expression analysis of mesenchymal stem cells derived from rat adipose tissue and bone marrow. Circ J. 2011;75:2260–8. doi: 10.1253/circj.CJ-11-0246.
    1. Khubutiya MS, Vagabov AV, Temnov AA, Sklifas AN. Paracrine mechanisms of proliferative, anti-apoptotic and anti-inflammatory effects of mesenchymal stromal cells in models of acute organ injury. Cytotherapy. 2014;16:579–85. doi: 10.1016/j.jcyt.2013.07.017.
    1. Xagorari A, Siotou E, Yiangou M, Tsolaki E, Bougiouklis D, Sakkas L, et al. Protective effect of mesenchymal stem cell-conditioned medium on hepatic cell apoptosis after acute liver injury. Int J Clin Exp Pathol. 2013;6:831–40.
    1. Liang X, Ding Y, Zhang Y, Tse HF, Lian Q. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant. 2014;23:1045–59. doi: 10.3727/096368913X667709.
    1. Elmadbouh I, Haider H, Jiang S, Idris NM, Lu G, Ashraf M. Ex vivo delivered stromal cell-derived factor-1alpha promotes stem cell homing and induces angiomyogenesis in the infarcted myocardium. J Mol Cell Cardiol. 2007;42:792–803. doi: 10.1016/j.yjmcc.2007.02.001.
    1. Lee JM, Jung J, Lee HJ, Jeong SJ, Cho KJ, Hwang SG, et al. Comparison of immunomodulatory effects of placenta mesenchymal stem cells with bone marrow and adipose mesenchymal stem cells. Int Immunopharmacol. 2012;13:219–24. doi: 10.1016/j.intimp.2012.03.024.

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