CellR4 2015; 3 (1): e1401

Understanding the Bioactivity of Stem Cells Seeded on Extracellular Matrix Scaffolds Produced from Discarded Human Kidneys: a Critical Step Towards a New Generation Bio-Artificial Kidney

Petrosyan A.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Orlando G.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
gorlando@wakehealth.edu
, Peloso A.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Wang Z.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Farney A. C.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Rogers G.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Katari R.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Da Sacco S.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Sedrakyan S.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, De Filippo R. E.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Stratta R. J.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Soker S.
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston Salem, NC, USA
, Perin L.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
lperin@chla.usc.edu

Topic: Regenerative Medicine   Category:

Abstract

Background: In the attempt to develop a clinically relevant platform for kidney bioengineering, we have previously demonstrated that human renal extracellular matrix scaffolds (hrECM) can be successfully and consistently produced from discarded kidneys. The hrECM are shown to maintain intricate architecture of macromolecules and vasculature. One of the main roadblocks in the bioengineering of transplantable kidneys is the incomplete understanding of how the extracellular matrix influences cellular behavior and response.

Materials and Methods: In the present study, we established a 3D in vitro model useful to investigate the interactions between stem cells and the extracellular matrix. Human amniotic fluid stem cells (hAFSC) were seeded on multiple samples of hrECM and cultured for up to 6 weeks. Histological examination of seeded scaffolds, analysis of secretomes, cell viability assessments, and PCR were carried out at different time points.

Results: We observed that hAFSC attach and proliferate when seeded on the matrix. hAFSC synthesize and secrete various chemokines and growth factors involved in angiogenesis (VEGF, TGF-alpha, IL-8) and inflammation, as well as mediators of matrix remodeling (MMP-2, TIMP-2). Ultrastructure analysis of cell-matrix interaction identified variation in cellular morphology based on renal compartment localization. Early kidney developmental markers, PAX and LIM1B, were expressed after 2 weeks of culture by hAFSC.

Conclusions: These results suggest that hrECM seeded with hAFSC behave as an effective and viable biosystem. Furthermore, this in vitro model may be used to study cell-matrix interactions and the mechanisms of tissue regeneration or repair occurring in the kidney.

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To cite this article

Petrosyan A.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Orlando G.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
gorlando@wakehealth.edu
, Peloso A.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Wang Z.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Farney A. C.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Rogers G.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Katari R.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Da Sacco S.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Sedrakyan S.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, De Filippo R. E.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
, Stratta R. J.
Department of General Surgery, Section of Transplantation, Wake Forest School of Medicine, Winston Salem, NC, USA
, Soker S.
Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston Salem, NC, USA
, Perin L.
GOFARR Laboratory, Saban Research Institute, Children’s Hospital, Department of Urology, University of Southern California, Los Angeles, CA, USA
lperin@chla.usc.edu
Understanding the Bioactivity of Stem Cells Seeded on Extracellular Matrix Scaffolds Produced from Discarded Human Kidneys: a Critical Step Towards a New Generation Bio-Artificial Kidney

CellR4 2015; 3 (1): e1401

Publication History

Published online: 05 Feb 2015