Lectin Histochemistry Showed a Heterogeneous Population of Cells Among Human Mesenchymal Stem Cells Isolated From Adipose Tissue
Objectives: Adipose tissue as an appropriate source of Mesenchymal Stem Cells (MSCs) has the potential to differentiate into multiple lineages. Glycoconjugates content of the MSCs can be considered as biomarkers in self-renewal, pluripotency and differentiation processes. In this study, the lectin profile of MSCs isolated from adipose tissue was detected and according to that, a subpopulation was determined.
Materials & Methods: MSCs were isolated from adipose tissue by explanting of the tissue pieces. The FITC-conjugated lectins, WGA, UEA, PNA, BSA and PWM were used to detect the terminal sugar residues. The cells were then counterstained with DAPI. The intensity of the reaction was evaluated by ImageJ software. The cells were also stained with PAS method.
Results: MSCs were reacted with all lectins with different intensity of the reactions. The cells reacted with WGA, UEA, and BSA “strongly” and with PWM “moderately” and with PNA with “weak” intensity. The morphological analysis of the isolated MSCs revealed the existence of the two different cell types in the cultures. Two types of cells were detected according to nucleus size and lectin reactivity. The cells with large nuclei constitute 20.62% of the total cells and stained significant more intensity by UEA and less intense with PWM (both P=0.014) and PNA (P=0.044). Flow cytometry with CD34 shows that these large cells were not endothelial cells.
Conclusion: The MSCs derived from adipose tissue seem to be a heterogeneous populations and lectin profile of the cells showed that they are different in the expression of the glycoconjugates.
Kern S, Eichler H, Stoeve J, Klüter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006; 24(5):1294–301. doi: 10.1634/stemcells.2005-0342
Dominici M, Paolucci P, Conte P, Horwitz EM. Heterogeneity ofmultipotent mesenchymal stromal cells: From stromal cells to stem cells and vice versa. Transplantation. 2009; 87:36–42. doi: 10.1097/tp.0b013e3181a283ee
Banas A, Teratani T, Yamamoto Y, Tokuhara M, Takeshita F, Quinn G, et al. Adipose tissue-derived mesenchymal stem cells as a source of human hepatocytes. Hepatology. 2007; 46(1):219–28. doi: 10.1002/hep.21704
Kawasaki-Oyama RS, Braile DM, Caldas HC, Leal JCF, Goloni-Bertollo EM, Pavarino-Bertelli ÉC, et al. [Blood mesenchymal stem cell culture from the umbilical cord with and without Ficoll-Paque density gradient method (Portuguese)]. Brazilian Journal of Cardiovascular Surgery. 2008; 23(1):29-34. PMID: 18719825
Talaei-Khozani T, Vojdani Z, Aliabadi E. A study of the glycoconjugates distribution of umbilical cord; a lectin histochemical study. Journal of Infertility and Reproductive Biology. 2013; 1(1):7-11.
Lavrov AV, Smirnikhina SA. Nuclear heterogeneity and proliferation activity of human adipose derived MSC-like cells. Cell and Tissue Biology. 2010; 4(5):452–6. doi: 10.1134/s1990519x1005007x
Baer PC, Geiger H. Adipose-derived mesenchymal stromal/stem cells: Tissue localization, characterization, and heterogeneity. Stem Cells International. 2012; 2012:1–11. doi: 10.1155/2012/812693
Mescher A. Junqueira’s basic histology: Text and atlas. New York: Mcgraw-hill; 2013.
Karsten U, Goletz S. What makes cancer stem cell markers different? Springer Plus. 2013; 2(1):301. doi: 10.1186/2193-1801-2-301
Yagi H, Kato K. Functional roles of glycoconjugates in the maintenance of stemness and differentiation process of neural stem cells. Glycoconjugate Journal. 2016; 1-7. doi: 10.1007/s10719-016-9707-x
Rosenman S, Gallatin W. Cell surface glycoconjugates in intercellular and cell-substratum interactions. Seminars in cancer biology. 1991; 2(6):357-66. PMID: 1810465
Sharon N. Lectins: Carbohydrate-specific reagents and biological recognition molecules. Journal of Biological Chemistry. 2007; 282(5):2753–64. doi: 10.1074/jbc.x600004200
Kang J, Park HM, Kim YW, Kim Y, Varghese S, Seok HK, et al. Control of mesenchymal stem cell phenotype and differentiation depending on cell adhesion mechanism. European Cells and Materials. 2014; 28:387–403. doi: 10.22203/ecm.v028a27
Talaei-Khozani T, Monsefi M, Ghasemi M. Lectins influence chondrogenesis and osteogenesis in limb bud mesenchymal cells. Glycoconjugate Journal. 2011; 28(2):89–98. doi: 10.1007/s10719-011-9326-5
Choi JH, Lyu SY, Lee HJ, Jung J, Park WB, Kim GJ. Korean mistletoe lectin regulates self-renewal of placenta-derived mesenchymal stem cells via autophagic mechanisms. Cell Proliferation. 2012; 45(5):420–9. doi: 10.1111/j.1365-2184.2012.00839.x
Diogo MM, Da Silva CL, Cabral JM. Separation technologies for stem cell bioprocessing. Biotechnology and Bioengineering. 2012; 109(11):2699-709. doi: 10.1002/bit.24706
Griesche N, Luttmann W, Luttmann A, Stammermann T, Geiger H, Baer PC. A simple modification of the separation method reduces heterogeneity of adipose-derived stem cells. Cells Tissues Organs. 2010; 192(2):106–15. doi: 10.1159/000289586
Rada T, Reis RL, Gomes ME. Distinct stem cells subpopulations isolated from human adipose tissue exhibit different chondrogenic and osteogenic differentiation potential. Stem Cell Reviews and Reports. 2010; 7(1):64–76. doi: 10.1007/s12015-010-9147-0
Anderson P, Carrillo-Gálvez AB, García-Pérez A, Cobo M, Martín F. CD105 (Endoglin)-negative murine mesenchymal stromal cells define a new multipotent subpopulation with distinct differentiation and im-munomodulatory capacities. PLoS ONE. 2013; 8(10):e76979. doi: 10.1371/journal.pone.0076979
Singh R. Peanut lectin stimulates proliferation of colon cancer cells by interaction with glycosylated CD44v6 isoforms and consequential activation of c-Met and MAPK: Functional implications for disease-associated glycosylation changes. Glycobiology. 2006; 16(7):594–601. doi: 10.1093/glycob/cwj108
Zayed S, Gaafar T, Samy R, Sabry D, Nasr A, Maksoud FA. Production of endothelial progenitor cells obtained from human Wharton's jelly using different culture conditions. Biotechnic & Histochemistry. 2016; 91(8):532-9. doi: 10.1080/10520295.2016.1250284
Song E, Lu CW, Fang LJ, Yang W. Culture and identification of endothelial progenitor cells from human umbilical cord blood. International Journal of Ophthalmology. 2010; 3(1): 49–53. doi: 10.3980/j.issn.2222-3959.2010.01.11
Zhou L, Xia J, Qiu X, Wang P, Jia R, Chen Y, et al. in vitro evaluation of endothelial progenitor cells from adipose tissue as potential angiogenic cell sources for bladder angiogen-esis. PLOS ONE. 2015; 10(2):0117644. doi: 10.1371/journal.pone.0117644
Mandai M, Ikeda H, Jin ZB, Iseki K, Ishigami C, Takahashi M. Use of lectins to enrich mouse es-derived retinal progenitor cells for the purpose of transplantation therapy. Cell Transplantation. 2010; 19(1):9–19. doi: org/10.3727/096368909x476599
Astori G, Vignati F, Bardelli S, Tubio M, Gola M, Albertini V, et al. “In vitro” and multicolor phenotypic characterization of cell subpopulations identified in fresh human adipose tissue stromal vascular fraction and in the derived mesenchymal stem cells. Journal of Translational Medicine. 2007; 5(1):55. doi: 10.1186/1479-5876-5-55
- There are currently no refbacks.
This work is licensed under a Creative Commons Attribution 3.0 License.
pISSN: 2423-5903 eISSN: 2538-4473