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Cell Biology JoVE (Open Access) Citable · DOI

Use of Human Perivascular Stem Cells for Bone Regeneration

DOI: 10.3791/2952-v
What you'll learn
  • ✓ Isolate human perivascular stem cells from adipose tissue via FACS
  • ✓ Implant PSC-scaffold constructs in three bone defect models
  • ✓ Evaluate bone regeneration outcomes in vivo
Protocol

Human perivascular stem cells (PSCs) are a novel stem cell class for skeletal tissue regeneration similar to mesenchymal stem cells (MSCs). PSCs can be isolated by FACS (fluorescence activated cell sorting) from adipose tissue procured during standard liposuction procedures, then combined with an osteoinductive scaffold to achieve bone formation in vivo.

Difficulty
advanced
Total time
~4–6 weeks per animal (including cell isolation, scaffold preparation, surgical implantation, and healing/analysis periods)
Model organism
Mouse (strain unspecified)
Biosafety
BSL-1

Steps

1
Implant PSC construct into muscle pouch

Create a muscle pouch surgical site and implant the perivascular stem cell-scaffold construct to assess bone formation in an ectopic location. This model serves as a baseline for osteoinductive capacity.

▶ 01:22
2
Implant PSC construct into calvarial defect

Create a critical-sized defect in the mouse calvarium (skull) and implant the PSC-scaffold construct to evaluate bone regeneration in an orthotopic cranial location.

▶ 02:24
3
Implant PSC construct into femoral segmental defect

Create a segmental bone defect in the femur and implant the PSC-scaffold construct to assess regeneration in a load-bearing long bone with high mechanical demands.

▶ 03:22
4
Analyze representative bone regeneration results

Present histological, radiographic, and quantitative outcome data from the three implantation models, demonstrating PSC efficacy in bone formation across different anatomical sites.

▶ 04:47
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