Ex vivo culture of peri-prostatic adipose tissue preserves local tissue architecture and maintains cell-cell interactions, crucial for studying prostate cancer biology.
This protocol enables the ex vivo culture and immortalisation of organ-specific adipocytes, providing a tool to study the mechanisms of obesity-driven diseases like prostate cancer.
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Obesity, officially recognised as a global epidemic by the World Health Organization, will soon overtake smoking as the largest preventable risk factor for cancer. By 2035, more than half the world’s population is expected to be overweight or obese with a significant increase in obesity-related health expenditures. However, despite the increase in prevalence and the overall lower life expectancy associated with obesity, mechanisms underpinning obesity-driven disease are not well understood. Adipocytes pose many challenges for in vitro culture due to their poor cell to surface attachment and low viability. Their large size and high lipid content can also present methodological challenges for downstream experiments. Several mouse and human-derived primary pre-adipocyte cell lines have been established over the years. However, they show limited renewal capacity and they cannot be cultured long term in vitro . Commercial cell lines available, which can be cultured long term, fail to represent organ-specific adipocyte heterogeneity. Adipose tissue from different organs and fat-depots can show significant heterogeneity in terms of metabolism, overall secretome and extracellular-matrix production. The prostate, for example, is surrounded by peri-prostatic adipose tissue (PPAT), volume of which is associated with increased risk of lethal prostate cancer and reduced therapy- response. Here we outline a protocol for ex vivo culture of fresh PPAT and non-prostatic adipose tissue (NPAT), which reflects donor and depot-specific characteristics. Ex vivo culture of PPAT/NPAT explants maintains cell-cell interactions and preserves local tissue architecture within adipose tissue. We have also described establishment of immortalised, patient PPAT-derived pre-adipocytes and patient-matched NPAT pre-adipocytes that can be in vitro- differentiated into mature adipocytes. The protocols outlined here could be readily adapted to other organ-specific fat depots such as mammary/bone marrow adipose tissue, and to tissues of non-human origin.
Grunberg et al. (Mon,) reported a other. Ex vivo culture of peri-prostatic adipose tissue preserves local tissue architecture and maintains cell-cell interactions, crucial for studying prostate cancer biology.