Home › Immunology › A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells
Immunology JoVE (Open Access) Citable · DOI

A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells

DOI: 10.3791/55841-v
What you'll learn
  • ✓ Isolate human kidney perivascular stromal cells
  • ✓ Perform whole organ perfusion with digestive enzymes
  • ✓ Enrich kPSCs using NG2-based selection
  • ✓ Culture and characterize clinical-grade kPSC populations
Protocol

Biopharma Insights Here we present a novel clinical grade isolation and culture method for kidney Perivascular Stromal Cells (kPSCs) based on whole organ perfusion with digestive enzymes and NG2-cell enrichment. With this method, it is possible to acquire sufficient cell numbers for cellular therapy.

Difficulty
advanced
Total time
~2-3 weeks (isolation, culture expansion, characterization)
Model organism
Human kidney tissue
Biosafety
BSL-2

Steps

1
Isolate kidney and perfuse with enzymes

Dissect kidney tissue and perform whole organ perfusion with digestive enzymes to release stromal cells.

▶ 00:53
2
Culture and expand kPSCs

Plate digested tissue and culture perivascular stromal cells under conditions supporting expansion.

▶ 03:48
3
Assess kPSC functional properties

Evaluate cell functionality using assays such as differentiation or immunomodulation.

▶ 05:46
4
Characterize human kPSC phenotype

Analyze representative human kPSC cultures for marker expression and morphology.

▶ 06:57
5
Summarize clinical-grade isolation method

Review key steps and outcomes of the clinical-grade kPSC isolation protocol.

▶ 07:49
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