TITLE:
Evaluation of TransFix® Mediated Stabilisation of Adipose-Derived Stromal Vascular Fraction for Delayed Flow Cytometry Analysis
AUTHORS:
Elisabeth Karsten, Judy Sung, Charlotte Morgan, Benjamin Herbert, Graham Vesey
KEYWORDS:
TransFix®, Adipose Tissue, Stromal Vascular Fraction, Immunophenotyping
JOURNAL NAME:
Open Journal of Regenerative Medicine,
Vol.3 No.3,
September
5,
2014
ABSTRACT: The increasing implementation
of multicentre studies has led to a need for the optimization of a method that allows
for accurate post-hoc analysis of patient biological samples. Assessment of total
cell number, viability and immunophenotype can present logistical challenges which
can be aided by batch processing. The increased sample storage time that this requires
necessitates the use of reagents to preserve cellular integrity, viability and immunophenotype.
TransFix is a stabilising reagent that has been developed for the preservation
of cell numbers and cell marker expression in peripheral whole blood for up to ten
days. This study investigated the use of TransFix reagent for the preservation
of the stromal vascular fraction (SVF) of collagenase digested adipose tissue. It
was demonstrated that TransFix was suitable for accurately measuring
nucleated SVF cell numbers for up to seven days as well as back calculating original
cell viability. It also stabilised three CD markers commonly used to identify populations
within SVF (CD90, CD31 and CD45) for up to seven days. There was no significant
difference between the number of CD90, CD31 and CD45 positive cells after stabilisation
at Day 7 compared to Day 0 unstabilised samples. The results suggest that TransFix can be used to preserve a biological mixed cell population from human adipose-derived
SVF for up to seven days for accurate post-hoc analysis.