Abstract
Progress in cardiac cell replacement therapies and tissue engineering critically depends on our ability to isolate functional cardiomyocytes (CMs) from heterogeneous cell mixtures. Label-free enrichment of cardiomyocytes is desirable for future clinical application of cell based products. Taking advantage of the physical properties of CMs, a microfluidic system was designed to separate CMs from neonatal rat heart tissue digest based on size using the principles of deterministic lateral displacement (DLD). For the first time, we demonstrate enrichment of functional CMs up to 91±2.4% directly from the digested heart tissue without any pre-treatment or labeling. Enriched cardiomyocytes remained viable after sorting and formed contractile cardiac patches in 3-dimensional culture. The broad significance of this work lies in demonstrating functional cell enrichment from the primary tissue digest leading directly to the creation of the engineered tissue.
Highlights
There are nearly 16 million people in North America alone suffering from coronary heart disease (CHD) with additional 770,000 new cases occurring annually contributing to $156 billion in medical expenses and lost productivity per year [1]
The cells focused by the sheath fluid flow through the sorting chamber (5 cm in length and 1.3 cm in width) and are collected in outlets 2–7 where outlet 2 is located in the middle of the sorting chamber
As cells move through the sorting chamber, large cells are expected to be displaced to the right side and collected in Outlets 3–7 while smaller cells are expected to stay in the middle of the chamber and collected in Outlet 2
Summary
There are nearly 16 million people in North America alone suffering from coronary heart disease (CHD) with additional 770,000 new cases occurring annually contributing to $156 billion in medical expenses and lost productivity per year [1]. During MI, a regional blockage in the coronary arteries constricts blood perfusion downstream which causes death of cardiomyocytes (CMs) in the infarct zone. Depending on the severity of infarction up to one billion CMs could be lost in the infarct zone [2]. These cells cannot be readily replaced by the heart since the adult mammalian CMs are considered to be terminally differentiated, have extremely low proliferation rates [3] and low turn-over rates in humans [4]. Cell replacement therapies offer the possibilities to develop new therapies for MI by replacing CMs lost during an MI either through cell injection or implantation of engineered cardiac patches
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