AMSC CD36+ subpopulations

Adipose-derived MSCs (AMSC) are currently utilized in clinical trials for various diseases including osteoarthritis, amyotrophic lateral sclerosis, and multiple system atrophy. Whilst AMSCs may be defined by the surface expression of classical markers including CD44, CD90, CD105, and absence of CD45, these cells may exhbit donor-to-donor differences in differentiation potential. In this study we evaluated the expression of CD36 amongst AMSCs and utilized magnetic cell sorting to enrich for CD36+ cells. Whole transcriptome RNA-sequencing was performed on unsorted, CD36+ enriched, and CD36+ depleted cells to determine gene expression difference between these populations of cells. CD36 sorted or unsorted cells were then plated and maintained in growth medium. Cells were expanded for 2 passages and passage 8 cells were plated for RNA-sequencing analysis. Trizol reagent was used to harvest cells when they were proliferating cells (~80%) or 100% confluent cells. AMSC donors 540 and 536 were expanded using clinical grade standard operating protocols and harvested for RNA-sequencing analysis upon confluence. AMSC donor 211 at passage 5 was harvested for magnetic cell sorting using CD36-APC antibody and anti-APC magnetic microbeads. Magnetic sorting was performed on these cells and the postive (enriched) and negative (depleted) fractions were collected. Additionally, a portion of the CD36-APC-microbead labelled cells but not sorted cells were kept as an unsorted population. CD36+ sorted fractions and unsorted cells then plated and expanded under growth conditions. After 2 passages, cells at passage 8 were plated for RNA-sequencing analysis, and samples were harvsted when they were either proliferating (~80% confluent) or 100% confluent.