J Korean Med Sci.  2015 Dec;30(12):1764-1776. 10.3346/jkms.2015.30.12.1764.

Advanced Properties of Urine Derived Stem Cells Compared to Adipose Tissue Derived Stem Cells in Terms of Cell Proliferation, Immune Modulation and Multi Differentiation

• Kang HS
• Choi SH
• Kim BS
• Choi JY
• Park GB
• Kwon TG
• Chun SY

• KMID: 2359960
• DOI: http://doi.org/10.3346/jkms.2015.30.12.1764

Abstract

Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.

Keyword

USCs; ADSCs; Stem Cell Characteristics; Multi-lineage Differentiation; Passages

MeSH Terms

Adult Stem Cells/*cytology/*immunology/transplantation
Biomarkers/metabolism
Cell Differentiation
Cell Lineage
Cell Proliferation
Cell Separation
Chromosomal Instability
Colony-Forming Units Assay
Humans
Karyotyping
Multipotent Stem Cells/cytology/immunology/transplantation
Subcutaneous Fat, Abdominal/*cytology
Transplantation, Autologous
Urine/*cytology
Biomarkers

Figure

• 

  Fig. 1 Comparisons of stem cell characters between USCs and ADSCs at passage 3, 5, and 7 (Representative images came from patient #91). (A) Cell morphology. Scale bars = 100 µm. (B) Cell proliferation analysis at day 1, 3, 5, 7, and 9. (C) Doubling time analysis. (D) Quantity of MSCs colonies. (E) Flow cytometric analysis for analysis of cell surface protein expression. USC, urine stem cell; ADSC, adipose tissue stem cell; P3, passage 3; P5, passage 5; P7, passage 7.

• 

  Fig. 2 Immune cell inhibitory effect of MSCs. PHA-activated peripheral blood mononuclear cells (PBMCs) was cultured by co-culture (A) or separate-culture (B), and the percent of inhibition efficiency was compared (C). USCs, urine stem cells; ADSCs, adipose tissue stem cells; PBMCs, peripheral blood mononuclear cells; P3, passage 3; P5, passage 5; P7, passage 7.

• 

  Fig. 3 G-banded karyotypes for USC and ADSC at passage 3, 5, and 7 (Representative images came from patient #91). Both USC and ADSC do not show any chromosomal aberrations. USC, urine stem cell; ADSC, adipose tissue stem cell; P3, passage 3; P5, passage 5; P7, passage 7.

• 

  Fig. 4 Comparisons of differentiation ability between USCs and ADSCs at passage 3, 5, and 7 (Representative immunocytochemical [ICC] images came from patient #91). (A) Stem cell markers expression of un-differentiated MSCs. (B) Myogenic differentiation potential analysis with MyoD gene expression and ICC. (C) Neurogenic differentiation potential analysis with β-Tubulin III gene expression and ICC. (D) Endothelial differentiation potential analysis with CD31 gene expression and ICC. (E) Adipogenic differentiation potential analysis with PPARγ gene expression and Oil-Red-O staining. (F) Osteogenic differentiation potential analysis with ALP gene expression and Alizarin-Red S staining. (G) Chondrogenic differentiation potential analysis with Aggrecan and Collagen type 4 gene expressions and Alcian blue staining. (H) Summarized table. Scale bars = 50 µm. USC, urine stem cell; ADSC, adipose tissue stem cell; differ USC, differentiated urine stem cell; differ ADSC, differentiated adipose tissue stem cell; P3, passage 3; P5, passage 5; P7, passage 7.

Reference

1. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002; 13:4279–4295.

2. Lenoir N. Europe confronts the embryonic stem cell research challenge. Science. 2000; 287:1425–1427.

3. Mehrabani D, Mehrabani G, Zare S, Manafi A. Adipose-derived stem cells (ADSC) and aesthetic surgery: a mini review. World J Plast Surg. 2013; 2:65–70.

4. Caplan AI. Mesenchymal stem cells. J Orthop Res. 1991; 9:641–650.

5. Choi YS, Noh SE, Lim SM, Lee CW, Kim CS, Im MW, Lee MH, Kim DI. Multipotency and growth characteristic of periosteum-derived progenitor cells for chondrogenic, osteogenic, and adipogenic differentiation. Biotechnol Lett. 2008; 30:593–601.

6. Song L, Young NJ, Webb NE, Tuan RS. Origin and characterization of multipotential mesenchymal stem cells derived from adult human trabecular bone. Stem Cells Dev. 2005; 14:712–721.

7. De Bari C, Dell'Accio F, Tylzanowski P, Luyten FP. Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum. 2001; 44:1928–1942.

8. Feng J, Mantesso A, Sharpe PT. Perivascular cells as mesenchymal stem cells. Expert Opin Biol Ther. 2010; 10:1441–1451.

9. Shi M, Ishikawa M, Kamei N, Nakasa T, Adachi N, Deie M, Asahara T, Ochi M. Acceleration of skeletal muscle regeneration in a rat skeletal muscle injury model by local injection of human peripheral blood-derived CD133-positive cells. Stem Cells. 2009; 27:949–960.

10. Dodson MV, Hausman GJ, Guan L, Du M, Rasmussen TP, Poulos SP, Mir P, Bergen WG, Fernyhough ME, McFarland DC, et al. Skeletal muscle stem cells from animals I. Basic cell biology. Int J Biol Sci. 2010; 6:465–474.

11. Belicchi M, Pisati F, Lopa R, Porretti L, Fortunato F, Sironi M, Scalamogna M, Parati EA, Bresolin N, Torrente Y. Human skin-derived stem cells migrate throughout forebrain and differentiate into astrocytes after injection into adult mouse brain. J Neurosci Res. 2004; 77:475–486.

12. Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, Young M, Robey PG, Wang CY, Shi S. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004; 364:149–155.

13. Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG, Shi S. SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci U S A. 2003; 100:5807–5812.

14. Baksh D, Yao R, Tuan RS. Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow. Stem Cells. 2007; 25:1384–1392.

15. Musina RA, Bekchanova ES, Sukhikh GT. Comparison of mesenchymal stem cells obtained from different human tissues. Bull Exp Biol Med. 2005; 139:504–509.

16. 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; 7:211–228.

17. Chun SY, Kim HT, Lee JS, Kim MJ, Kim BS, Kim BW, Kwon TG. Characterization of urine-derived cells from upper urinary tract in patients with bladder cancer. Urology. 2012; 79:1186.e1–1186.e7.

18. Zuk PA. Tissue engineering craniofacial defects with adult stem cells? Are we ready yet? Pediatr Res. 2008; 63:478–486.

19. Marino G, Rosso F, Cafiero G, Tortora C, Moraci M, Barbarisi M, Barbarisi A. Beta-tricalcium phosphate 3D scaffold promote alone osteogenic differentiation of human adipose stem cells: in vitro study. J Mater Sci Mater Med. 2010; 21:353–363.

20. Zhang Y, McNeill E, Tian H, Soker S, Andersson KE, Yoo JJ, Atala A. Urine derived cells are a potential source for urological tissue reconstruction. J Urol. 2008; 180:2226–2233.

21. Zhu Y, Liu T, Song K, Fan X, Ma X, Cui Z. Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochem Funct. 2008; 26:664–675.

22. Srikanth G. What are the functional passages that can be used for mesenchymal stem cells? accessed on 5 September 2014. Available at http://www.researchgate.net/post/.

23. Zhou T, Benda C, Duzinger S, Huang Y, Li X, Li Y, Guo X, Cao G, Chen S, Hao L, et al. Generation of induced pluripotent stem cells from urine. J Am Soc Nephrol. 2011; 22:1221–1228.

24. Hematti P. Mesenchymal stromal cells and fibroblasts: a case of mistaken identity? Cytotherapy. 2012; 14:516–521.

25. Haasters F, Prall WC, Anz D, Bourquin C, Pautke C, Endres S, Mutschler W, Docheva D, Schieker M. Morphological and immunocytochemical characteristics indicate the yield of early progenitors and represent a quality control for human mesenchymal stem cell culturing. J Anat. 2009; 214:759–767.

26. Aguilera A, Loureiro J, Gónzalez-Mateo G, Selgas R, López-Cabrera M. The mesothelial to mesenchymal transition a pathogenic and therapeutic key for peritoneal membrane failure. In : Aguilera A, editor. The latest in peritoneal dialysis. Rijeka, Croatia: InTech Press;2013.

27. Mark P, Kleinsorge M, Gaebel R, Lux CA, Toelk A, Pittermann E, David R, Steinhoff G, Ma N. Human mesenchymal stem cells display reduced expression of CD105 after culture in serum-free medium. Stem Cells Int. 2013; 2013:698076.

28. 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; 347:419–427.

29. Le Blanc K, Tammik C, Rosendahl K, Zetterberg E, Ringdén O. HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol. 2003; 31:890–896.

30. Hass R, Kasper C, Böhm S, Jacobs R. Different populations and sources of human mesenchymal stem cells (MSC): a comparison of adult and neonatal tissue-derived MSC. Cell Commun Signal. 2011; 9:12.

31. Vilchez D, Boyer L, Morantte I, Lutz M, Merkwirth C, Joyce D, Spencer B, Page L, Masliah E, Berggren WT, et al. Increased proteasome activity in human embryonic stem cells is regulated by PSMD11. Nature. 2012; 489:304–308.

32. Fong CY, Peh GS, Gauthaman K, Bongso A. Separation of SSEA-4 and TRA-1-60 labelled undifferentiated human embryonic stem cells from a heterogeneous cell population using magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS). Stem Cell Rev. 2009; 5:72–80.

33. Trubiani O, Zalzal SF, Paganelli R, Marchisio M, Giancola R, Pizzicannella J, Bühring HJ, Piattelli M, Caputi S, Nanci A. Expression profile of the embryonic markers nanog, OCT-4, SSEA-1, SSEA-4, and frizzled-9 receptor in human periodontal ligament mesenchymal stem cells. J Cell Physiol. 2010; 225:123–131.

34. Ohyama M. Hair follicle bulge: a fascinating reservoir of epithelial stem cells. J Dermatol Sci. 2007; 46:81–89.

35. Amoh Y, Li L, Katsuoka K, Penman S, Hoffman RM. Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. Proc Natl Acad Sci U S A. 2005; 102:5530–5534.

36. Xu ZC, Zhang Q, Li H. Differentiation of human hair follicle stem cells into endothelial cells induced by vascular endothelial and basic fibroblast growth factors. Mol Med Rep. 2014; 9:204–210.

37. Wobus AM, Boheler KR. Stem cells. Berlin: Springer;2008.