Abstract 14009: Human Patient-Specific SKP-Derived Vascular Smooth Muscle Cells Carry Signatures of Type II Diabetes
Background: Vascular complications of type-II diabetes (T2D) are a major cause of morbidity and mortality. We previously showed that skin-derived precursors (SKP) from foreskin and lip of children can be differentiated at ~90% efficiency into vascular smooth muscle cells (VSMC), and that SKP-derived VSMC remain responsive to β-adrenergic agonists and integrate into functioning vessels in vivo. Objectives: To determine if patient-specific VSMC can be derived from SKP found in chest or leg incisions of aged adults, and to assess whether they could represent a platform for studying T2D.
Methods & Results: SKP derived from aged patients with or without T2D undergoing cardiothoracic surgery were differentiated into VSMC at similar frequencies to foreskin-derived SKP from children (>80% yield, as evidenced by ASMA+/Calponin+/Smoothelin-B+ confocal immunofluorescence). However, adult patient-specific SKP failed to passage, suggesting accelerated senescence and/or disease. Although SKP from diabetics were also able to differentiate into VSMC, they were isolated at significantly lower efficiencies compared to non-diabetics (2.49x105±0.06 vs. 5.96x105 ±0.09 cells/g skin, P<0.05). By contrast, no differences in SKP isolation efficiency were observed between male vs. female, or chest vs. leg incisions. The EC50 of phenylephrine (PE)-induced VSMC responses were significantly higher for VSMC derived from adult- vs. child SKP (3015±2566 nM vs. 2.2±1.2 nM, P<0.05). By contrast, VSMC derived from diabetics demonstrated lower EC50 values than non-diabetics (129±77 nM vs. 3015±2566 nM). Further differences in Ca2+ handling characteristics were observed in VSMC derived from SKP obtained from adults with T2D as compared to those derived from non-diabetics, supporting the notion of a disease-specific phenotype. Summary: SKP derived from aged adults fail to passage and are less abundant in T2D. Adult patient-specific SKP-derived VSMC exhibit properties of aged cells and are functionally less responsive to β-adrenergic agonists than SKP-derived VSMC from children. VSMC from adults with T2D exhibited increased responsiveness to PE compared to non-diabetics suggesting a disease (diabetes)-specific phenotype.
- © 2013 by American Heart Association, Inc.