BackgroundCell 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.MethodsThis 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.ResultsLarge 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.ConclusionAlthough 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.
Pre-analytical enrichment of cell populations prior to therapeutic delivery is of paramount interest throughout the fields of regenerative medicine and clinical interventional therapies. Enrichment of a cell population typically involves two aspects: (a) the increase in concentration of particular subpopulation of the total cell fraction by means of removal of other cells of no interest to the particular interrogation; and (b) improvement of resolution of signal by removal of 'noise' mostly arising from cellular debris in the treated sample. In this research, leukocyte populations were obtained from erythrocyte-depleted primary whole blood from human adults and subjected to flow through acoustic fields within the ultrasound range to remove cellular debris. It was possible to demonstrate aggregation and holding of leukocytes by using ultrasound within the frequency range 11.448-11.483 MHz, which facilitated removal of cellular debris by washing under continuous perfusion. The T-lymphocyte population were phenotypically characterized using CD4/CD8 (T(h)/T(c)) immunocytochemistry by flow cytometry and demonstrated a significant decrease in 'false-positive' events during cellular analysis, due to the efficient eradication of non-specifically reactive cells and tissue debris from the cell populations of interest. Therefore, it was possible to conclude that flow through an ultrasonic acoustic system was capable of providing a non-destructive method for the hyper-purification of primary derived cell populations, with potential exploitations throughout the fields of cellular research, medical diagnostics and clinical therapies.
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