Stem cellMesenchymal Stem Cells Delivered at the Subcapsule of the Kidney Ameliorate Renal Disease in the Rat Remnant Kidney Model
Section snippets
MSC Isolation and Expansion
Briefly, bone marrow from femur and tibia of male Wistar rats (180–220 g, obtained from local colony) were flushed out and cultivated with Dulbecco's modified Eagle's medium–low glucose (DMEM-low) supplemented with 10% fetal bovine serum. MSC were isolated using their characteristic to adhere to plastic culture dishes. After 2 weeks of culture, adherent cells were harvested by trypsinization, washed with phosphate-buffered saline (PBS), and kept on ice until the moment of the infusion. For the
MSC Culture and Characterization
MSC isolated from femur and tibia bone marrow formed colonies of fibroblast-like cells after about 12–20 days of culture, and grew into a homogeneous population of cells, with a mainly fusiform morphology. Analysis of cell surface markers using flow cytometry showed that MSC were positive for CD44 (61 ± 8%) and CD90 (85 ± 4%), and mostly negative for CD31 (2.5 ± 0.5%), CD34 (4 ± 1%), and CD45 (19 ± 4%; Fig 1.).
Osteogenic Differentiation of MSC
After 3 weeks of culture in osteogenic induction medium, MSC acquired an
Discussion
In this study, we have shown that MSC isolated from rat bone marrow and injected into the subcapsule of remnant kidney rats with the 5/6 Nx experimental model of chronic progressive nephropathy significantly ameliorated renal function 30 days after nephrectomy, however, it was already detected at 15 days.
Although no unique surface marker can be used for specific characterization of MSC, these cells can be identified by combined expression of cell surface markers and exclusion of others. In our
Acknowledgments
The authors thank Laila Casado and Fernanda Cobucci for their skillful technical assistance and gratefully acknowledge Dr Denis Feliers for his excellent help in reviewing the manuscript.
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Supported by CNPq (Brazilian Council for Scientific and Technologic Development) number 552644/2005-6 and by FAPESP (Foundation for Research Support) number 2006/56628-4.