Arrayed cellular microenvironments for identifying culture and differentiation conditions for stem, primary and rare cell populations

Abstract

During the development of an organism, cells are exposed to a myriad of signals, structural components and scaffolds, which collectively make up the cellular microenvironment. The majority of current developmental biology studies examine the effect of individual or small subsets of molecules and parameters on cellular behavior, and they consequently fail to explore the complexity of factors to which cells are exposed. Here we describe a technology, referred to as arrayed cellular microenvironments (ACMEs), that allows for a high-throughput examination of the effects of multiple extracellular components in a combinatorial manner on any cell type of interest. We will specifically focus on the application of this technology to human pluripotent stem cells (hPSCs), a population of cells with tremendous therapeutic potential, and one for which growth and differentiation conditions are poorly characterized and far from defined and optimized. A standard ACME screen uses the technologies previously applied to the manufacture and analysis of DNA microarrays, requires standard cell-culture facilities and can be performed from beginning to end within 5-10 days.

A chemically defined substrate for the expansion and neuronal differentiation of human pluripotent stem cell-derived neural progenitor cells

“…Cells are globally seeded onto the arrays and due to the non-fouling nature of the acrylamide hydrogel cells only adhere to sites of polymer deposition. As we have previously shown (Brafman et al, 2010;Brafman et al, 2012;Brafman et al, 2009a;Brafman et al, 2013;Brafman et al, 2009b), paracrine signaling between neighboring spots is limited and each spot can be treated an independent 'well'. Combined with high-content automated microscopy which allows for quantitative single-cell analysis, this technology can be used to screen the effect of thousands of unique polymer compositions on the fate of any cell type of interest on a single microscope slide.…”

“…Arrays of polymers were fabricated as previously described (Brafman et al, 2010;Brafman et al, 2012). Briefly, glass slides were cleaned, silanized, and then functionalized with a polyacrylamide gel layer.…”

High‐throughput investigation of endothelial‐to‐mesenchymal transformation (EndMT) with combinatorial cellular microarrays

“…In addition, the microarray platforms were a great tool to study stem cell differentiation in the combinatorial effect of microenvironments (Anderson et al, 2004;Brafman et al, 2012;Dolatshahi-Pirouz et al, 2014;Flaim et al, 2005;Gobaa et al, 2011;Klim et al, 2010;Mei et al, 2010;Soen et al, 2006). These platforms enabled the identification of cell regulating microenvironmental parameters that would have been difficult to find with conventional cell culture methods (Charnley et al, 2009) and significantly reduce the preparation and optimization process of new biomaterials (Dolatshahi-Pirouz et al, 2011).…”

Infrared spectroscopy as a new tool for studying single living cells: Is there a niche?

“…For these reasons, the study of single cells is important to investigate processes that cannot be otherwise well understood in heterogeneous cell samples [67]. In addition, the isolation and identification of single cells for biomolecular assays requires defined protocols and strict techniques [10]. The spectroscopic analysis of single cells can be considered an alternative interesting approach, even if not all the spectroscopic tools are suitable for this study.…”