Skip to main content

Advertisement

SpringerLink
Log in

HEK293-Conditioned Medium Altered the Expression of Renal Markers WT1, CD2AP, and CDH16 in the Human Adipose Mesenchymal Stem Cells

  • Original Research
  • Published:
Regenerative Engineering and Translational Medicine Aims and scope Submit manuscript

Abstract

The conditioned medium (CM) could be used as the suitable source of growth factors in the regenerative medicine which had the ability to enhance efficiency of stem cells in the repair of injured tissues. In the present study, we studied the effect of conditioned medium derived from HEK293 (HEK293-CM) on the expression of CD2AP, CDH16, and WT1 genes in the human adipose-mesenchymal stem cells (AD-MSCs). For this purpose, the human AD-MSCs were treated with different concentrations (5%, 10%, or 15%) of prepared HEK293-CM. Then, the expression level of three renal markers, CD2AP, CDH16, and WT1, was determined in the treated cells as compared with untreated cells as the control group. Our data showed that treatment with 15%CM-5%FBS-24 h and 5%CM-10%FBS-72 h was significantly associated with enhanced expression of CD2AP and WT1 in AD-MSCs (p < 0.05). Furthermore, increased expression of CDH16 and CD2AP was observed in AD-MSCs treated by 10%CM-1%FBS-48 h (p = 0.0002 and p = 0.02 for CDH16 and CD2AP, respectively). The results obtained from this study indicated that HEK293-CM had the ability to increase the characteristics related to the renal cell lineage in AD-MSCs. This approach could be introduced as the novel method to enhance the ability of stem cells in the repair of renal injuries.

Lay Summary

Different studies showed that the conditioned medium provided the novel approach to enhance the stem cells efficiency in the repair of the injured tissues. The growth factors secreted from the cells into the medium could be used as the alternative option to stimulate the differentiation ability of stem cell. In the present study, we prepared the conditioned medium from HEK293 (HEK293-CM) and evaluated its effect on the adipose-mesenchymal stem cells (AD-MSCs).The obtained results indicated that HEK293-CM had the ability to increase the characteristics related to the renal cell lineage in AD-MSCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Batchelder CA, Lee CC, Matsell DG, Yoder MC, Tarantal AF. Renal ontogeny in the rhesus monkey (Macaca mulatta) and directed differentiation of human embryonic stem cells towards kidney precursors. Differentiation; research in biological diversity. 2009;78(1):45–56.

    Article  CAS  Google Scholar 

  2. - Kang, M., and Han, Y. M. (2014) Differentiation of human pluripotent stem cells into nephron progenitor cells in a serum and feeder free system. PloS One 9(4): e94888.

  3. Lange C, Tögel F, Ittrich H, Clayton F, Nolte-Ernsting C, Zander AR, Westenfelder C. Administered mesenchymal stem cells enhance recovery from ischemia/reperfusion-induced acute renal failure in rats. Kidney Int. 2005;68(4):1613–7.

    Article  Google Scholar 

  4. Bi B, Schmitt R, Israilova M, Nishio H, Cantley LG. Stromal cells protect against acute tubular injury via an endocrine effect. J Am Soc Nephrol. 2007;18(9):2486–96.

    Article  Google Scholar 

  5. - Morigi, M., Rota, C., and Remuzzi, G. (2016) Mesenchymal stem cells in kidney repair. Methods in molecular biology (Clifton, N.J.) 1416: 89–107.

  6. Villanueva S, González F, Lorca E, Tapia A, López VG, Strodthoff R, Fajre F, Carreño JE, Valjalo R, Vergara C, Lecanda M, Bartolucci J, Figueroa FE, Khoury M. Adipose tissue-derived mesenchymal stromal cells for treating chronic kidney disease: a pilot study assessing safety and clinical feasibility. Kidney research and clinical practice. 2019;38(2):176–85.

    Article  Google Scholar 

  7. Roushandeh AM, Bahadori M, Roudkenar MH. Mesenchymal stem cell-based therapy as a new horizon for kidney injuries. Arch Med Res. 2017;48(2):133–46.

    Article  CAS  Google Scholar 

  8. Pittenger MF, Discher DE, Péault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. NPJ Regenerative medicine. 2019;4:22.

    Article  Google Scholar 

  9. Bochon B, Kozubska M, Surygała G, Witkowska A, Kuźniewicz R, Grzeszczak W, Wystrychowski G. Mesenchymal stem cells-potential applications in kidney diseases. Int J Mol Sci. 2019;20(10):2462.

    Article  Google Scholar 

  10. Shi H, Patschan D, Dietz GP, Bähr M, Plotkin M, Goligorsky MS. Glial cell line-derived neurotrophic growth factor increases motility and survival of cultured mesenchymal stem cells and ameliorates acute kidney injury. Am J Physiol Renal Physiol. 2008;294(1):F229–35.

    Article  CAS  Google Scholar 

  11. Zhong S, He X, Li Y, Lou X. Conditioned medium enhances osteogenic differentiation of induced pluripotent stem cell-derived mesenchymal stem cells. Tissue Eng Regen Med. 2019;16(2):141–50.

    Article  CAS  Google Scholar 

  12. Sun X, Cheng L, Duan H, Lu G. Effects of an endothelial cell-conditioned medium on the hematopoietic and endothelial differentiation of embryonic stem cells. Cell Biol Int. 2009;33(11):1201–5.

    Article  CAS  Google Scholar 

  13. - Ardeshirylajimi, A., Vakilian, S., Salehi, M., and Mossahebi-Mohammadi, M. (2017) Renal differentiation of mesenchymal stem cells seeded on nanofibrous scaffolds improved by human renal tubular cell lines-conditioned medium. ASAIO J Am Soc Art Int Org 1992, 63(3), 356–363.

  14. Ashokkumar B, Vaziri ND, Said HM. Thiamin uptake by the human-derived renal epithelial (HEK-293) cells: cellular and molecular mechanisms. Am J Physiol Renal Physiol. 2006;291(4):F796–805.

    Article  CAS  Google Scholar 

  15. Torban E, Goodyer PR. Effects of PAX2 expression in a human fetal kidney (HEK293) cell line. Biochem Biophys Acta. 1998;1401(1):53–62.

    Article  CAS  Google Scholar 

  16. Lin YC, Boone M, Meuris L, Lemmens I, Van Roy N, Soete A, Reumers J, Moisse M, Plaisance S, Drmanac R, Chen J, Speleman F, Lambrechts D, Van de Peer Y, Tavernier J, Callewaert N. Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations. Nat Commun. 2014;5:4767.

    Article  CAS  Google Scholar 

  17. - Mohammadi, B., Esmaeilizadeh, Z., Rajabibazl, M., Ghaderian, S.M.H., Omrani, M.D., Fazeli, Z. (2020) Preconditioning of human adipose tissue-derived mesenchymal stem cells with HEK293-coditioned media can influence on the expression of BMP2, BMP6 and BMP11: Potential application in the treatment of renal lesions. Gene Rep 21; 100912.

  18. Brzoska M, Geiger H, Gauer S, Baer P. Epithelial differentiation of human adipose tissue-derived adult stem cells. Biochem Biophys Res Commun. 2005;330(1):142–50.

    Article  CAS  Google Scholar 

  19. - Pawitan J. A. (2014) Prospect of stem cell conditioned medium in regenerative medicine. BioMed Res Int 2014: 965849.

  20. Vizoso FJ, Eiro N, Cid S, Schneider J, Perez-Fernandez R. Mesenchymal stem cell secretome: toward cell-free therapeutic strategies in regenerative medicine. Int J Mol Sci. 2017;18(9):1852.

    Article  Google Scholar 

  21. Chen Q, Liang Q, Zhuang W, Zhou J, Zhang B, Xu P, Ju Y, Morita Y, Luo Q, Song G. Tenocyte proliferation and migration promoted by rat bone marrow mesenchymal stem cell-derived conditioned medium. Biotechnol Lett. 2018;40(1):215–24.

    Article  CAS  Google Scholar 

  22. Shimode K, Iwasaki N, Majima T, Funakoshi T, Sawaguchi N, Onodera T, Minami A. Bone marrow stromal cells act as feeder cells for tendon fibroblasts through soluble factors. Tissue Eng. 2007;13(2):333–41.

    Article  CAS  Google Scholar 

  23. Wang Y, He G, Guo Y, Tang H, Shi Y, Bian X, Zhu M, Kang X, Zhou M, Lyu J, Yang M, Mu M, Lai F, Lu K, Chen W, Zhou B, Zhang J, Tang K. Exosomes from tendon stem cells promote injury tendon healing through balancing synthesis and degradation of the tendon extracellular matrix. J Cell Mol Med. 2019;23(8):5475–85.

    Article  CAS  Google Scholar 

  24. Karavanova ID, Dove LF, Resau JH, Perantoni AO. Conditioned medium from a rat ureteric bud cell line in combination with bFGF induces complete differentiation of isolated metanephric mesenchyme. Development (Cambridge, England). 1996;122(12):4159–67.

    Article  CAS  Google Scholar 

  25. Li CY, Wu XY, Tong JB, Yang XX, Zhao JL, Zheng QF, Zhao GB, Ma ZJ. Comparative analysis of human mesenchymal stem cells from bone marrow and adipose tissue under xeno-free conditions for cell therapy. Stem Cell Res Ther. 2015;6(1):55.

    Article  Google Scholar 

  26. Shih NY, Li J, Cotran R, Mundel P, Miner JH, Shaw AS. CD2AP localizes to the slit diaphragm and binds to nephrin via a novel C-terminal domain. Am J Pathol. 2001;159(6):2303–8.

    Article  CAS  Google Scholar 

  27. Kawachi H, Fukusumi Y. New insight into podocyte slit diaphragm, a therapeutic target of proteinuria. Clin Exp Nephrol. 2020;24(3):193–204.

    Article  Google Scholar 

  28. - Shih, N. Y., Li, J., Karpitskii, V., Nguyen, A., Dustin, M. L., Kanagawa, O., Miner, J. H., and Shaw, A. S. (1999) Congenital nephrotic syndrome in mice lacking CD2-associated protein. Science (New York, N.Y.) 286(5438): 312–315.

  29. Grunkemeyer JA, Kwoh C, Huber TB, Shaw AS. CD2-associated protein (CD2AP) expression in podocytes rescues lethality of CD2AP deficiency. J Biol Chem. 2005;280(33):29677–81.

    Article  CAS  Google Scholar 

  30. Asanuma K, Campbell KN, Kim K, Faul C, Mundel P. Nuclear relocation of the nephrin and CD2AP-binding protein dendrin promotes apoptosis of podocytes. Proc Natl Acad Sci USA. 2007;104(24):10134–9.

    Article  CAS  Google Scholar 

  31. Schiffer M, Mundel P, Shaw AS, Böttinger EP. A novel role for the adaptor molecule CD2-associated protein in transforming growth factor-beta-induced apoptosis. J Biol Chem. 2004;279(35):37004–12.

    Article  CAS  Google Scholar 

  32. Chen, Y., Chen, J., Wan, J., Gao, N., Cui, J., You, D., and Zou, Z. (2017) Bone marrow-derived mesenchymal stem cells ameliorate nephrosis through repair of impaired podocytes. Clinical and investigative medicine. Medecine clinique et experimentale 40(1): E13–E24.

  33. Zoja C, Garcia PB, Rota C, Conti S, Gagliardini E, Corna D, Zanchi C, Bigini P, Benigni A, Remuzzi G, Morigi M. Mesenchymal stem cell therapy promotes renal repair by limiting glomerular podocyte and progenitor cell dysfunction in adriamycin-induced nephropathy. Am J Physiol Renal Physiol. 2012;303(9):F1370–81.

    Article  CAS  Google Scholar 

  34. Dong L, Pietsch S, Englert C. Towards an understanding of kidney diseases associated with WT1 mutations. Kidney Int. 2015;88(4):684–90.

    Article  CAS  Google Scholar 

  35. Guo JK, Menke AL, Gubler MC, Clarke AR, Harrison D, Hammes A, Hastie ND, Schedl A. WT1 is a key regulator of podocyte function: reduced expression levels cause crescentic glomerulonephritis and mesangial sclerosis. Hum Mol Genet. 2002;11(6):651–9.

    Article  CAS  Google Scholar 

  36. Hosono S, Luo X, Hyink DP, Schnapp LM, Wilson PD, Burrow CR, Reddy JC, Atweh GF, Licht JD. WT1 expression induces features of renal epithelial differentiation in mesenchymal fibroblasts. Oncogene. 1999;18(2):417–27.

    Article  CAS  Google Scholar 

  37. Thedieck C, Kuczyk M, Klingel K, Steiert I, Müller CA, Klein G. Expression of Ksp-cadherin during kidney development and in renal cell carcinoma. Br J Cancer. 2005;92(11):2010–7.

    Article  CAS  Google Scholar 

  38. Singaravelu K, Padanilam BJ. In vitro differentiation of MSC into cells with a renal tubular epithelial-like phenotype. Ren Fail. 2009;31(6):492–502.

    Article  CAS  Google Scholar 

  39. Nonnis S, Maffioli E, Zanotti L, Santagata F, Negri A, Viola A, Elliman S, Tedeschi G. Effect of fetal bovine serum in culture media on MS analysis of mesenchymal stromal cells secretome. EuPA Open Proteom. 2016;10:28–30.

    Article  CAS  Google Scholar 

Download references

Funding

The present study was financially supported by the “Research Department of the School of Medicine Shahid Beheshti University of Medical Sciences” (Grant No 11196). The present study was approved by the Ethics Committee of the School of Medicine Shahid Beheshti University of Medical Sciences (Tehran, Iran; Ethical code: IR.SBMU. MSP.REC.1397. 6).

Author information

Authors and Affiliations

  1. Department of Medical Genetics, Faculty of Medicine, Shahid Beheshti University of Medical Sciences,, Tehran, Iran

    Zahra Fazeli & Zahra Esmaeilizadeh

  2. Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mir Davood Omrani & Sayyed Mohammad Hossein Ghaderian

  3. Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Masoumeh Rajabibazl

Authors
  1. Zahra Fazeli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zahra Esmaeilizadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mir Davood Omrani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sayyed Mohammad Hossein Ghaderian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masoumeh Rajabibazl
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Z. Fazeli supervised the project and wrote the manuscript; Z. Esmaeilizadeh carried out the experiments; MD. Omrani and SMH. Ghaderian contributed to the interpretation of the results; M Rajabibazl helped in writing the manuscript.

Corresponding author

Correspondence to Zahra Fazeli.

Ethics declarations

Consent to Participate

This study was an in vitro study and it did not require providing the informed consent.

Consent to Publish

All authors read and confirmed the final manuscript.

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fazeli, Z., Esmaeilizadeh, Z., Omrani, M.D. et al. HEK293-Conditioned Medium Altered the Expression of Renal Markers WT1, CD2AP, and CDH16 in the Human Adipose Mesenchymal Stem Cells. Regen. Eng. Transl. Med. 8, 456–462 (2022). https://doi.org/10.1007/s40883-021-00246-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40883-021-00246-7

Keywords

Search

Navigation