Urine-Derived Stem Cells: Differentiation Potential into Smooth-Muscle Cells and Urothelial Cell

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Abstract


Background: Tissue engineering of low urinary tract organs requires biopsy of urinary bladder material. The current study describes non-invasive approach of obtaining autologous stem cells from urine of healthy adults. These cells were studied for potential to differentiate into epithelial cells and smooth muscle cells of the urinary bladder. Aims: To describe properties of urine-derived stem cells (USCs) and investigate their differentiation potential for tissue engineering of low urinary tract organs. Materials and Methods: USCs were isolated from urine of healthy volunteers with centrifugation and seeded in media to 24-well plates. Expression of stem cells markers (CD73, CD90, CD105, CD34, CD45, CD29, CD44, CD54, SSEA4) by USCs was assessed with flow cytometry. Expression of specific markers of smooth muscle cells and urothelial cells was assessed with fluorescence microscopy with following computational image analysis. Results: Median number of USCs per 100 ml urine was 6. Doubling time for USC was 1.44±0.528 days (n=4) and there were 26.3±4.79 population doublings for USC cultures (n=4). Median expression of markers of postnatal stem cells was CD73 ― 79.8%, CD90 ― 56.6%, CD105 ― 40.7%, CD34 <1.0%, CD45 <2.0%, CD29 >99.0%, CD44 >99.0%, CD54 ― 97.7% and SSEA4 >99.0%. Treatment of cells with high concentration of EGF in media with low concentration of FBS for 10 days increased cytokeratin (CK) expression to 24.9% for CK AE1/AE3 and to 7.6% for CK 7. Treatment of USCs with media inducing smooth muscle differentiation for 10 days increased expression of α-smooth muscle actin to 79.6% and expression of calponin to 97.6%. Conclusions: USCs are cells that can be found in urine in small quantities. They have high proliferative potential and express markers of postnatal stem cells. Under effect of PDGF-BB and TGF-β1 they differentiate into smooth muscle cells.


Igor A. Vasyutin

I.M. Sechenov First Moscow State Medical University (Sechenov University)

Author for correspondence.
Email: ivasyutin@yahoo.com
ORCID iD: 0000-0003-0594-7423

Russian Federation

MD.

8 bld 2, Trubetskaya street, 119991 Moscow.

SPIN-код: 1872-8347

Aleksey V. Lyundup

I.M. Sechenov First Moscow State Medical University (Sechenov University)

Email: lyundup@gmail.com
ORCID iD: 0000-0002-0102-5491

Russian Federation

MD, PhD.

8 bld 2, Trubetskaya street, 119991 Moscow.

SPIN-код: 4954-3004

Sergey L. Kuznetsov

I.M. Sechenov First Moscow State Medical University (Sechenov University)

Email: vakmedbiol@rambler.ru
ORCID iD: 0000-0002-0704-1660

Russian Federation

MD, PhD, Professor.

8 bld 2, Trubetskaya street, 119991 Moscow.

SPIN-код: 3824-2646

  1. Lee YJ, Kim SW. Current management of urethral stricture. Korean J Urol. 2013;54(9):561–569. doi: 10.4111/kju.2013.54.9.561.2.
  2. Atala A, Kasper FK, Mikos AG. Engineering complex tissues. Sci Transl Med. 2012;4(160):160rv12. doi: 10.1126/scitranslmed.3004890.3.
  3. Vasyutin I, Zerihun L, Ivan C, Atala A. Bladder organoids and spheroids: potential tools for normal and diseased tissue modelling. Anticancer Res. 2019;39(3):1105–1118. doi: 10.21873/anticanres.13219.4.
  4. Васютин И.А., Люндуп А.В., Винаров А.З., и др. Реконструкция уретры с помощью технологий тканевой инженерии // Вестник РАМН. ― 2017. ― Т.72. ― №1. ― С. 17−25. doi: 10.15690/vramn771.
  5. Orabi H, AbouShwareb T, Zhang Y, et al. Cell-seeded tubularized scaffolds for reconstruction of long urethral defects: a preclinical study. Eur Urol. 2013;63(3):531–538. doi: 10.1016/j.eururo.2012.07.041.6.
  6. Atala A, Danilevskiy M, Lyundup A, et al. The potential role of tissue-engineered urethral substitution: clinical and preclinical studies. J Tissue Eng Regen Med. 2017;11(1):3–19. doi: 10.1002/term.2112.7.
  7. Bhargava S, Patterson JM, Inman RD, et al. Tissue-engineered buccal mucosa urethroplasty-clinical outcomes. Eur Urol. 2008;53(6):1263–1269. doi: 10.1016/j.eururo.2008.01.061.8.
  8. Yang B, Zheng JH, Zhang YY. Myogenic differentiation of mesenchymal stem cells for muscle regeneration in urinary tract. Chin Med J (Engl). 2013;126(15):2952–2959. doi: 10.5772/55961.
  9. Osborn SL, Thangappan R, Luria A, et al. Induction of human embryonic and induced pluripotent stem cells into urothelium. Stem Cells Transl Med. 2014;3(5):610–619. doi: 10.5966/sctm.2013-0131.10.
  10. Wang Z, Wen Y, Li YH, et al. Smooth muscle precursor cells derived from human pluripotent stem cells for treatment of stress urinary incontinence. Stem Cells Dev. 2016;25(6):453–461. doi: 10.1089/scd.2015.0343.11.
  11. Kanematsu A, Yamamoto S, Iwai-Kanai E, et al. Induction of smooth muscle cell-like phenotype in marrow-derived cells among regenerating urinary bladder smooth muscle cells. Am J Pathol. 2005;166(2):565–573. doi: 10.1016/S0002-9440(10)62278-X.
  12. Zhang Y, McNeill E, Tian H, et al. Urine derived cells are a potential source for urological tissue reconstruction. J Urol. 2008;180(5):2226–2233. doi: 10.1016/j.juro.2008.07.023.14.
  13. Bharadwaj S, Liu G, Shi Y, et al. Multipotential differentiation of human urine-derived stem cells: potential for therapeutic applications in urology. Stem Cells. 2013;31(9):1840–1856. doi: 10.1002/stem.1424.15.
  14. Shi JG, Fu WJ, Wang XX, et al. Transdifferentiation of human adipose-derived stem cells into urothelial cells: potential for urinary tract tissue engineering. Cell Tissue Res. 2012;347(3):737–746. doi: 10.1007/s00441-011-1317-0.

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