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Abstract: SA-PO012

Isolation and Purification of Mesangial Cells for Bioengineered Kidneys

Session Information

Category: Bioengineering

  • 400 Bioengineering

Authors

  • Stachel, MacKenna A., Miromatric Medical Inc., Eden Prarie, Minnesota, United States
  • Nelson, Tori, Miromatric Medical Inc., Eden Prarie, Minnesota, United States
  • Beck, Emily C., Miromatric Medical Inc., Eden Prarie, Minnesota, United States
Background

With over 85,000 patients waiting for a kidney transplant, bioengineered kidneys (BEKs) offer a way to address this large clinical need. BEKs are created from isolated native cell types from donor kidneys not accepted for transplant. Of these isolated cell types, mesangials help maintain the glomerular filtration barrier of the kidney. Here, an isolation and purification process are presented to isolate mesangial populations that can be used for glomerular recapitulation of the BEK.

Methods

Using enzymatic digestion, whole glomeruli were isolated from human kidneys and then plated on tissue culture flasks for 14 days. Mesangials were purified from the outgrowths using CD140b/PDGFRβ positive magnetic bead sorting. Purification of the bound mesangial cell population was confirmed using flow cytometry. Post sort, mesangials were expanded, then cryo-banked at passage 2. Cryopreserved cells were thawed and expanded to passage 5, where the mesangials were seeded into a BEK along with other native glomerular cell types.

Results

The bound mesangial population isolated from 6 kidney donors had 89.2% ± 13.4% CD140b positive cells. To assess the potential of isolated mesangials for cryo-banking and expansion, mesangials were cryopreserved, and expanded in 2D up through passage 5. The mesangials remained viable through 2D culture (95.3% ± 2.9% at passage 5, n=7), and were seeded and cultured in 3D in a BEK (n=1) along with other glomerular cell types (podocytes). This BEK demonstrated improved protein and RBC filtration in comparison to BEKs seeded only with podocytes and parietal epithelial cells, after 60 minutes of whole porcine blood perfusion.

Conclusion

Mesangial cells are a necessary cell type known for their structural support as well as their contractile properties to aid in ultrafiltration. This study demonstrates a process in which the mesangial population can be isolated from human donor kidneys, purified, cryo-banked, and expanded for making BEKs. Furthermore, when seeded into a BEK, the mesangials demonstrated improved BEK filtration function. Cryo-banked populations of mesangial cells, as well as other cell types, will allow for BEK creation on demand, thus helping to reduce the number of patients waiting for a kidney transplant.

Funding

  • Commercial Support – Miromatrix Medical Inc.