Jorge Castillo’s bio

My name is Jorge Castillo, I am originally from Ecuador. I completed my Bachelor of Science at the University of Toronto. I am now a first-year Master of Science student in the laboratory of Dr. Ian Rogers at Mount Sinai Hospital in Toronto. My project involves generating an ex-vivo model of kidney disease utilizing a bioreactor designed to house an organ and maintain it for multiple days. I am excited about regenerative medicine approaches to treating disease and the different ways that we can use human and non-human sources to regenerate functional tissue. In my free time I enjoy spending time with friends and playing soccer.

Developing an Ex-vivo organ perfusion system to model kidney disease

Anupama Bhadwal, Alex Cho, Ian Rogers

Chronic kidney disease (CKD) affects 11-13% of the population worldwide. Disease models that aid in uncovering the disease pathogenesis and developing novel therapeutics are needed. Multiple animal models of CKD involve invasive procedures that can affect the animal’s health and experiment results. Also, since the affected organs cannot be analyzed without invasive techniques disease stages are missed. Ex-vivo whole-organ perfusion (EVOP) provides a solution since it enables real time, longitudinal analysis at the whole-organ level and the practical administration of treatments. Also, controlled culture conditions lead to reduced alterations between experiments. We hypothesize that a mouse kidney can be maintained using EVOP for 14 days. Following kidney isolation, renal artery, and vein cannulation, the kidney was cultured using EVOP for 4, 7, and 14 days.
We determined that tissue viability along with proper expression of key kidney proteins such as nephrin, LTL, AQP2 were maintained for up to 7 days. We determined that our system meets the oxygen requirement of a mouse kidney. By analyzing the urine generated by EVOP kidneys, we demonstrated normal renal functioning as the kidney was able to reabsorb proteins found in the culture medium and maintain normal urine glucose levels. However, cell death was found at day 7.

Future steps include optimizing the culture medium to extend EVOP times to 14 days. We aim to enhance the growth conditions by supplementing the culture medium with essential and non-essential amino acids and fatty acids to supply the metabolic needs of the kidney. We aim to implement Flolan, a known vasodilator used in whole organ ex-vivo perfusion studies. Further, we will also analyze gene expression to help improve the EVOP model.