Novel fluid shear‐based dissociation device for improved single cell dissociation of spheroids and cell aggregates

Abstract

Biological industries commonly rely on bioreactor systems for the large-scale production of cells. Cell aggregation, clumping, and spheroid morphology of certain suspension cells make their large-scale culture challenging. Growing stem cells as spheroids is indispensable to retain their stemness, but large spheroids (>500 µm diameter) suffer from poor oxygen and nutrient diffusion, ultimately resulting in premature cell death in the centers of the spheroids. Despite this, most large-scale bioprocesses do not have an efficient method for dissociating cells into single cells, but rely on costly enzymatic dissociation techniques. Therefore, we tested a proof-of-concept fluid shear-based mechanical dissociator that was designed to dissociate stem cell spheroids and aggregates. Our prototype was able to dissociate cells while retaining high viability and low levels of apoptosis. The dissociator also did not impact long-term cell growth or spheroid formation. Thus, the dissociator introduced here has the potential to replace traditional dissociation methods. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:293-298, 2018.

Microfluidic Devices in the Fast‐Growing Domain of Single‐Cell Analysis

“…Because, biological systems need a large‐scale production of single‐cells, while in‐addition to hydrolytic shear forces, their system relied on an expensive enzymatic dissociation technique. Triantafillu et al . presented a fluid shear‐based mechanical dissociator (Figure e).…”

“…( e) Schematic of FSS driven tissue dissociation.Reprinted with permission from Ref. [18].Copyright 2017 Wiley-VCH. processing is still ac hallenge.…”

“…processing is still ac hallenge. Because, biological systems need al arge-scalep roduction of single-cells, while in-addition to hydrolytic shear forces, their system relied on an expensive enzymatic dissociation technique.T riantafillu et al [18] presented a fluid shear-based mechanical dissociator ( Figure 2e). The prototype consists of as yringe pump, as yringe, and PEEK tubing with inner dimeter of 125 and 254 mma nd lengthso f4 6a nd 96 cm.…”

“…The mechanical dissociation method is of particular interest because in this technique the isolated single‐cells maintain the endogenous surface marker expression. These devices offer easier scale‐up and higher productivity, yet the design for more practical application are demanding. Future devices must be able to process a real human specimen with the ability to separate the heterogeneous cells population in the specimen.…”