Directed differentiation of human mesenchymal stem cells toward a cardiomyogenic fate commitment through formation of cell aggregates

Abstract

Cell morphology is known to modulate the multipotential lineage commitment of stem cells. We provide a new strategy to induce the early lineage commitment of human mesenchymal stem cells (hMSCs) toward a cardiomyogenic fate through the formation of cell aggregates. A surface-immobilized polyamidoamine dendrimer with fifth generation of dendron structure was used during the culturing of hMSCs. These hMSCs cultured on the G5 surface formed aggregates through active migration and division. More than 22% of cardiac troponin-T (cTnT)-positive (cTnT +) cells in aggregates formed on the dendrimer surface; the population formed on the dendrimer surface was higher than that in conventional culture vessel. When cell aggregate were reseeded onto a fresh G5 surface, single cells migrated out of the aggregates, proliferated, and formed new aggregates. This passage method, accompanied with repetitive aggregate dispersion and formation, was applied to cultures over 40 days. The proportion of cTnT + cells increased to 62% by the end of third passage. Our results suggest that culturing hMSCs on G5 surface results in directed commitment of the hMSCs toward a cardiomyocyte-like fate.

Migration-driven aggregate behaviors of human mesenchymal stem cells on a dendrimer-immobilized surface direct differentiation toward a cardiomyogenic fate commitment

“…Dynamic changes in b-catenin expression and transcriptional activity were regulated in response to altered cell adhesion during migration. Consistent with previous results (15), cell aggregates on the G5 surface induced cTnT expression due to the morphological changes, and the results from this study suggest that b-catenin signaling in response to changes in migration behaviors of hMSC aggregates may be implicated in the guidance of cardiomyogenic fate. bCatenin was expressed at the interface among cells in the aggregates on the G5 surface and the low-binding culture surface.…”

“…Migration-driven aggregate behaviors direct differentiation toward a cardiomyogenic fate commitment through cellecell dissociation The spontaneous formation and migratory behaviors of aggregates affect cell-substrate and cellecell adhesions within aggregates and thereby regulate cell differentiation signaling (14,15). Within migrating aggregates, cellecell and cellsubstrate adhesions are integrated so that cells maintain interactions with neighboring cells and the underlying substratum.…”

“…Quantitative assessment of cardiomyogenic differentiation was conducted as described previously (15). Briefly, cells incubated for a specified number of days were collected by enzymatic treatment and re-seeded on the PS surface.…”

“…Repetitive collapse and re-formation of the aggregates increased the population of cardiomyocyte-like cells by passage culture of aggregates on a new dendrimer surface (15). In this study, we investigated the migratory behaviors of hMSC aggregates on the dendrimer surface to enhance directed differentiation toward a cardiomyogenic lineage.…”

Designing a blueprint for next‐generation stem cell bioprocessing development

“… Design of cell behavior: the behavior of stem cells such as cell–cell interactions, cell‐substrate interactions, and cell migration plays an important role in the regulation of cell fate decisions (Guilak et al, ; Kim & Kino‐oka, ; Kim & Kino‐oka, ). Considering the fact that development processes contain multiple branchpoints, many studies have reported a cell culture strategy for inducing direct differentiation and producing a more homogeneous population of pluripotent cells (Kim, Ogawa, Yamada, Taya, & Kino‐oka, ; Kim, Sugawara, Fujinaga, & Kino‐oka, ; Ogawa, Kim, & Kino‐oka, ). This will require regulation of strategies pertaining to multiple branchpoints during development processes, namely: (a) the deviation points that arise from undifferentiated states; (b) the first lineage branchpoints in the developmental path for directing the differentiation of cells toward a specified lineage; and (c) the branch point sequences for favoring or restricting development toward specialized cell types that comprise a tissue.…”