Tissue dissociation miniaturized platform for uterine stem cell isolation and culture

Endometrial stem cells provide a readily accessible source of adult stem cells suitable for various stem cell based therapies. The first essential step in the process of stem cell isolation, sorting and culture is tissue dissociation into single cell suspension. Tissue dissociation efficiency determines the number of resulting viable cells which in turn could affect the feasibility of the subsequent stem cell application. Microfluidic based platforms could provide several advantages over current conventional tissue dissociation techniques (e.g. mechanical or enzymatic or both) and help the optimization of the process towards a better reproducible cellular outcome. Objective: To develop a simple miniaturized platform for endometrial tissue dissociation which can be used to evaluate and optimize experimental conditions necessary for producing single cell suspension with minimal cell damage. Methods: A Poly-DiMethylSiloxane (PDMS) channel microfluidic platform was designed. The platform includes a tissue dissociation chamber where raw endometrial tissue was incubated. Trypsin was delivered through an inlet channel while another inlet enabled parallel delivery of a cell suspension medium and an outlet channel received the downstream dissociated endometrial cells. A cell size based filtration step is designed to trap large debris using two rows of posts. A viability stain is used to test the viability of cells produced at different enzyme-tissue contact durations. Conclusion: We present a simple microfluidics based platform for endometrial tissue dissociation that can be used for subsequent endometrial stem cell research applications and can be further applied to other tissues.