Pino 2023 Abstract IOC162

Pino MF, Kapar SG, Laurent NL, Erickson ML, Sparks LM (2023) Characterization of human subcutaneous adipose-derived stem cells isolated from healthy young and old individual. Mitochondr Physiol Network 28.02

Link: IOC162

Pino Maria F, Kapar Sophie G, Laurent Nicolas L, Erickson Melissa L, Sparks Lauren M (2023)

Event: IOC162

Adipose tissue, which is the crucial energy reservoir and endocrine organ for the maintenance of systemic glucose, lipid, and energy homeostasis, undergoes significant changes during aging. Ageing also impacts the circadian clock (CC) machinery of peripheral organs including white adipose tissue. Together, these alterations cause age-related disease in the elderly population. The aim of this study is to investigate the transcriptional expression of circadian clock genes, cell proliferation rates and mitochondrial capacity of adipose-derived stem cells (ASCs) isolated from abdominal subcutaneous white adipose tissue (scWAT) from old and young individuals.

10 old (5 females) and 10 young individuals (5 females) participated in this study. To measure CC related genes, proliferating ASCs were synchronized with 30% fetal bovine serum (FBS) for two hours, then media was replaced with 2% FBS and RNA was collected every six hours for a duration of 48 hrs and targeted gene expression was measured by qRT-PCR. To measure cells proliferation, ASCs were plated in 96 well plate and cell proliferation was measured with CellTiter-Glow luminescent cell viability kit (Promega, USA) for a period of 72 hrs. Mitochondrial capacity, oxygen consumption rates, were measured in proliferating ASCs via high-resolution respirometry using the Oxygraph-2K (Oroboros instruments, Innsbruck, Austria).

An increase in mRNA levels of CLOCK and PER2 and a loss of rhythmicity for CLOCK and CRY1 were observed in ASCs from older individuals compared to the young. mRNA levels and rhythmicity of BMAL, PER1, DBP, NR1D1 and NR1D2 were not altered by age. We are currently determining cell proliferation and mitochondrial capacity in ASCs. Overall, age does not seem to affect CC rhythmicity in ASCs isolated from scWAT from old and young individuals.

• Bioblast editor: Plangger M • O2k-Network Lab: US FL Orlando Sparks LM

Labels: MiParea: Respiration 

HRR: Oxygraph-2k 

Affiliations

Translational Research Institute for Metabolism and Diabetes, Florida Hospital, Orlando, USA.