Clonal analysis of individual human embryonic stem cell differentiation patterns in microfluidic cultures

Abstract

Heterogeneity in the clonal outputs of individual human embryonic stem cells (hESCs) confounds analysis of their properties in studies of bulk populations and how to manipulate them for clinical applications. To circumvent this problem we developed a microfluidic device that supports the robust generation of colonies derived from single ESCs. This microfluidic system contains 160 individually addressable chambers equipped for perfusion culture of individual hESCs that could be shown to match the growth rates, marker expression and colony morphologies obtained in conventional cultures. Use of this microfluidic device to analyze the clonal growth kinetics of multiple individual hESCs induced to differentiation revealed variable shifts in the growth rate, area per cell and expression of OCT4 in the progeny of individual hESCs. Interestingly, low OCT4 expression, a slower growth rate and low nuclear to cytoplasmic ratios were found to be correlated responses. This study demonstrates how microfluidic systems can be used to enable large scale live-cell imaging of isolated hESCs exposed to changing culture conditions, to examine how different aspects of their variable responses are correlated.

Microfluidic Single-Cell Manipulation and Analysis: Methods and Applications

“…Sikorski et al developed a microfluidic device that supports the robust generation of colonies derived from single human embryonic stem cells (hESCs) [103]. The use of this device to analyze the clonal growth of CAIS hESCs demonstrated its ability to reveal the heterogeneity of differentiation patterns displayed by clonally tracked hESC.…”

Developments in cell culture systems for human pluripotent stem cells

“…Microfluidic chips were designed for analysis of hPSC response to treatments as single colonies[111,112]. Similar systems have been used to understand how growth factor signals could impact cell fate determination[113,114], comparable to organ-on-a-chip platforms for cancer and somatic cells. Microfluidic devices are powerful tools for research in cellular function, cell fate determination as well as disease modeling.…”

Cancer Liquid Biopsy Using Integrated Microfluidic Exosome Analysis Platforms

“…It is an emerging interdisciplinary technology based on concepts derived from fluid mechanics and other diverse engineering fields (e.g., materials, microelectronics, biomedical engineering, etc.). [ 18 ] Microfluidic devices, commonly referred to as microfluidic chips, Lab‐on‐a‐Chip (LOC), and micro total analysis system (μTAS) devices, can integrate multi‐step functionality onto a single chip, including cell culture, [ 19 ] sample pretreatment, [ 20 ] separation, [ 21 ] and detection [ 22 ] to conserve samples and achieve rapid processing with high efficiency. [ 23,24 ] Over the past two decades, microfluidic technology has been successfully applied to selective sorting [ 25,26 ] and detection [ 27,28 ] of various types of cells.…”