Elsevier

Cytotherapy

Volume 15, Issue 6, June 2013, Pages 663-672
Cytotherapy

Original paper
Mesenchymal stromal cells
Bone marrow-derived mesenchymal stromal cells from patients with end-stage renal disease are suitable for autologous therapy

https://doi.org/10.1016/j.jcyt.2013.01.010Get rights and content

Abstract

Background aims

Mesenchymal stromal cells (MSCs) are pluripotent cells that have immunosuppressive and reparative properties in vitro and in vivo. Although autologous bone marrow (BM)-derived MSCs are already clinically tested in transplant recipients, it is unclear whether these BM cells are affected by renal disease. We assessed whether renal failure affected the function and therapeutic potential of BM-MSCs.

Methods

MSCs from 10 adults with end-stage renal disease (ESRD) and 10 age-matched healthy controls were expanded from BM aspirates and tested for phenotype and functionality in vitro.

Results

MSCs from ESRD patients were >90% positive for CD73, CD90 and CD105 and negative for CD34 and CD45 and showed a similar morphology and differentiation capacity as MSCs from healthy controls. Of importance for their clinical utility, growth characteristics were similar in both groups, and sufficient numbers of MSCs were obtained within 4 weeks. Messenger RNA expression levels of self-renewal genes and factors involved in repair and inflammation were also comparable between both groups. Likewise, microRNA expression profiling showed a broad overlap between ESRD and healthy donor MSCs. ESRD MSCs displayed the same immunosuppressive capacities as healthy control MSCs, demonstrated by a similar dose-dependent inhibition of peripheral blood mononuclear cell proliferation, similar inhibition of proinflammatory cytokines tumor necrosis factor-α and interferon-γ production and a concomitant increase in the production of interleukin-10.

Conclusions

Expanded BM-MSCs procured from ESRD patients and healthy controls are both phenotypically and functionally similar. These findings are important for the potential autologous clinical application of BM-MSCs in transplant recipients.

Introduction

It has been postulated in recent years that mesenchymal stromal cells (MSCs) may be useful in modifying and potentially reversing loss of function in kidney disease and in preventing allograft rejection in transplant recipients 1, 2, 3, 4, 5, 6. MSCs were first discovered in bone marrow (BM) aspirates but have also been isolated and expanded from various other adult tissues (7). At the present time, no unique phenotype has been identified that allows the prospective isolation of MSCs. Their isolation and characterization relies primarily on their ability to adhere to plastic, their multi-lineage differentiation and their membrane antigen profile (8).

Pre-clinical studies indicate that administration of MSCs ameliorates renal injury and accelerates tissue repair, and numerous experimental studies have demonstrated beneficial effects after solid organ transplantation 2, 4, 5, 9, 10, 11, 12, 13. The potential mechanism of MSC-induced kidney repair has been addressed in numerous studies 10, 13, 14, 15, 16, 17, 18. There is increasing evidence that the process of trans-differentiation is rare and of limited biologic relevance (15). It is currently believed that paracrine factors are responsible for the effects of MSCs, including their mitogenic, anti-apoptotic, angiogenic and various immune-modulating properties 19, 20, 21, 22, 23, 24, 25.

In human kidney transplantation, there is great interest in the therapeutic application of MSCs, and the first clinical trials using autologous BM-MSCs in kidney transplantation with the initial focus on their potential as induction therapy and to treat allograft rejection have started 20, 26, 27. A concern for the use of allogeneic MSCs is the potential induction of an allo-response with negative consequences for MSC efficacy and potentially for graft survival in the case of transplantation. A concern for the use of autologous MSCs includes their potential dysfunction secondary to the underlying disease, as reported in systemic lupus erythematosus, immune thrombocytopenic purpura, rheumatoid arthritis, multiple myeloma and aplastic anemia 28, 29, 30, 31, 32, and a potential safety risk related to genetic stability. A few pre-clinical studies have reported on the impact of renal disease on the phenotype and function of MSCs. In mice, functional incompetence of MSCs was reported after exposure to uremic conditions (33). In human BM-MSCs, uremic serum induced an osteoblast-like phenotype accompanied by matrix remodeling and calcification (34), whereas human adipose tissue-derived MSCs are not affected by renal disease (35). None of these studies has studied human BM-MSCs isolated from patients with end-stage renal disease (ESRD); so far, BM-MSCs are the only cell product used in clinical trials with MSCs in nephrology.

We performed a head-to-head comparison between BM-MSCs from patients with ESRD and from healthy controls regarding their phenotypic and functional characteristics. More recently, microRNAs (miRNAs) have been shown to play a critical role in differentiation and MSC functionality (36), and their dysregulation has been shown in various diseases including kidney diseases 37, 38. Because miRNA expression in MSCs from patients with ESRD is completely unknown, we have included in this study comparison of miRNA expression in MSCs from patients with ESRD and healthy controls. We believe our results to be important for clinical application in transplant recipients.

Section snippets

Patients and controls

BM samples were collected from patients with ESRD and from healthy controls that were either BM donors in a clinical renal transplantation setting or patients undergoing orthopedic surgery. Ethics committee approval was obtained for the aspiration protocol, and individual written consents were obtained from all individuals. Two age-matched groups of 10 individuals each were compiled for the analyses.

Isolation and culture of MSCs

Heparinized BM was aspirated under local or general anesthesia. The mononucleated cell fraction

Isolation, expansion and characterization of MSCs

Because differentiation potential and proliferation rate are reported to decline in aged human MSCs 39, 40, we selected two age-matched study populations and used expanded MSC populations at similar low passage numbers (P3–4). Healthy controls (five women and five men) had a median age of 64 years (±7.98), which was comparable to the patients with ESRD. Patient demographics are presented in Table I. The MSC cultures of patients with ESRD and healthy controls showed similar expansion kinetics

Discussion

The great potential for MSCs as a therapeutic tool for allograft rejection after renal transplantation is supported by positive results in different experimental models 5, 9, 11, 13, 41. In these studies, recipient and donor-derived MSCs were shown to have better immunomodulatory properties than third-party MSCs 41, 42, 43, 44. However, donor-derived MSCs were also reported to cause sensitization, and application with autologous MSCs seems preferable 45, 46, 47. The first clinical trials using

Acknowledgments

We thank Ms E. Steeneveld and A. Loor for collecting and archiving donor and patient material and Ms E. van Beelen for the multiplex cytokine analysis. This study was sponsored by the Dutch Organisation for Sciences (NWO/ZonMW; TAS and Veni), a Nephrosearch Grant and the European Community's Seventh Framework Programme (FP7/2007–13, HEALTH-F5-2008-223007 STAR-T REK).

Author contributions are as follows: MEJR, research design, performance of research and data analysis and manuscript preparation;

References (53)

  • M. Roemeling-van Rhijn et al.

    Mesenchymal stem cells derived from adipose tissue are not affected by renal disease

    Kidney Int

    (2012)
  • K. Chandrasekaran et al.

    Role of microRNAs in kidney homeostasis and disease

    Kidney Int

    (2012)
  • C. Fehrer et al.

    Mesenchymal stem cell aging

    Exp Gerontol

    (2005)
  • K. Stenderup et al.

    Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells

    Bone

    (2003)
  • F.C. Popp et al.

    Mesenchymal stem cells can induce long-term acceptance of solid organ allografts in synergy with low-dose mycophenolate

    Transpl Immunol

    (2008)
  • H.P. Zhou et al.

    Administration of donor-derived mesenchymal stem cells can prolong the survival of rat cardiac allograft

    Transplant Proc

    (2006)
  • W. Ge et al.

    Infusion of mesenchymal stem cells and rapamycin synergize to attenuate alloimmune responses and promote cardiac allograft tolerance

    Am J Transplant

    (2009)
  • A.J. Nauta et al.

    Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting

    Blood

    (2006)
  • N. Eliopoulos et al.

    Allogeneic marrow stromal cells are immune rejected by MHC class I- and class II-mismatched recipient mice

    Blood

    (2005)
  • B.D. Humphreys et al.

    Mesenchymal stem cells in acute kidney injury

    Annu Rev Med

    (2008)
  • M. Morigi et al.

    Human bone marrow mesenchymal stem cells accelerate recovery of acute renal injury and prolong survival in mice

    Stem Cells

    (2008)
  • Casiraghi F, Noris M, Remuzzi G. Immunomodulatory effects of mesenchymal stromal cells in solid organ transplantation....
  • M.E. Reinders et al.

    Multipotent mesenchymal stromal cell therapy in renal disease and kidney transplantation

    Nephrol Dial Transplant

    (2010)
  • M. Morigi et al.

    Mesenchymal stem cells are renotropic, helping to repair the kidney and improve function in acute renal failure

    J Am Soc Nephrol

    (2004)
  • M. Franquesa et al.

    Mesenchymal stem cell therapy prevents interstitial fibrosis and tubular atrophy in a rat kidney allograft model

    Stem Cells Dev

    (2012)
  • M. Franquesa et al.

    The impact of mesenchymal stem cell therapy in transplant rejection and tolerance

    Curr Opin Organ Transplant

    (2012)
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