High-efficiency cellular reprogramming with microfluidics

Abstract

We report that the efficiency of reprogramming human somatic cells to induced pluripotent stem cells (hiPSCs) can be dramatically improved in a microfluidic environment. Microliter-volume confinement resulted in a 50-fold increase in efficiency over traditional reprogramming by delivery of synthetic mRNAs encoding transcription factors. In these small volumes, extracellular components of the TGF-β and other signaling pathways exhibited temporal regulation that appears critical to acquisition of pluripotency. The high quality and purity of the resulting hiPSCs (μ-hiPSCs) allowed direct differentiation into functional hepatocyte- and cardiomyocyte-like cells in the same platform without additional expansion.

Monitoring cell secretions on microfluidic chips using solid-phase extraction with mass spectrometry

“…PDMS valves have demonstrated great versatility for on-chip fluid handling, from automating fluid flow over select cell chambers [44] to transporting cells via peristaltic pumping [45]. Pneumatically-actuated valves are integrated on PDMS chips through multilayer assembly [46].…”

Ductular reaction-on-a-chip: Microfluidic co-cultures to study stem cell fate selection during liver injury

“…Several reports, including from our group, have demonstrated that microfluidic channels may enhance cell function by helping to accumulate cell-secreted signals182223. This was shown to be true for stem cells18, cancer cells24, and most recently for hepatocytes34.…”

“…This is being attributed to enhanced accumulation of endogenous growth factors and autocrine signals in confined volumes of microfluidic chambers operating under diffusion dominant transport conditions (low flow regime). Such enhanced autocrine signals have been observed in stem cells182223, cancer cells24 and in primary hepatocytes1925.…”

Universal Microfluidic System for Analysis and Control of Cell Dynamics

“…To accomplish this, precise control of environmental parameters such as cell density, surface properties, fluidic exchange, media and growth factor delivery, and humidity is required. Microfluidic devices using membrane valves allow automated generation of well-controlled programmable conditions mimicking in vivo microenvironments (Gomez-Sjoberg et al, 2007;Lecault et al, 2011;Luni et al, 2016;Tay et al, 2010). Despite the advances in integrated microfluidics, however, long-term cultivation of sensitive cell types, such as primary mammalian cells, remains challenging.…”