Evaluation of a novel non-destructive catch and release technology for harvesting autologous adult stem cells

PLoS One. 2013;8(1):e53933. doi: 10.1371/journal.pone.0053933. Epub 2013 Jan 22.

Abstract

Background: Cell based therapies are required now to meet the critical care needs of paediatrics and healthy ageing in an increasingly long-lived human population. Repair of compromised tissue by supporting autologous regeneration is a life changing objective uniting the fields of medical science and engineering. Adipose stem cells (adSCs) are a compelling candidate for use in cell based medicine due to their plasticity and residence in numerous tissues. Adipose found in all animals contains a relatively high concentration of stem cells and is easily isolated by a minimally invasive clinical intervention; such as liposuction.

Methods: This study utilised primary rat adipose to validate a novel strategy for selecting adult stem cells. Experiments explored the use of large, very dense cell-specific antibody loaded isolation beads (diameter 5x-10x greater than target cells) which overcome the problem of endocytosis and have proved to be very effective in cell isolation from minimally processed primary tissue. The technique also benefited from pH mediated release, which enabled elution of captured cells using a simple pH shift.

Results: Large beads successfully captured and released adSCs from rat adipose, which were characterised using a combination of microscopy, flow cytometry and PCR. The resultant purified cell population retains minimal capture artefact facilitating autologous reperfusion or application in in vitro models.

Conclusion: Although evidenced here for adSCs, this approach provides a technological advance at a platform level; whereby it can be applied to isolate any cell population for which there is a characterised surface antigen.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology*
  • Adipose Tissue / metabolism
  • Adult
  • Adult Stem Cells / cytology*
  • Adult Stem Cells / metabolism
  • Animals
  • Cell Count
  • Cell Separation / methods*
  • Flow Cytometry
  • Humans
  • Microscopy, Fluorescence
  • Rats
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Thy-1 Antigens / genetics
  • Thy-1 Antigens / metabolism

Substances

  • Thy-1 Antigens

Grants and funding

This research was funded by Strategic Projects to Access Research and Knowledge (SPARK), United Kingdom (grant number not available)(SPARK is a specific funding call offered by Knowledge Transfer Network) United Kingdom). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.