EMAIL THIS PAGE TO A FRIEND

Biomaterials

The use of agarose microwells for scalable embryoid body formation and cardiac differentiation of human and murine pluripotent stem cells.


PMID 23332176

Abstract

In most pluripotent stem cell differentiation protocols the formation of embryoid bodies (EBs) is an important step. Here we describe a rapid, straightforward soft lithography approach for the preparation of hydrophilic silicon masters from different templates and the subsequent production of patterned agarose-DMEM microwell surfaces for scalable well standardized stem cell aggregation and EB formation. The non-adhesive agarose microwell plates represent an accurate replication of the templates' topography and were used for aggregation of murine induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs). Direct microscopic assessment by time-lapse analysis demonstrated rapid formation of uniformly shaped EBs from murine iPSCs with similar or even more consistent results concerning size distribution and harvesting efficiency compared to the commonly used but time-consuming hanging drop technique. For human ESCs, homogenous aggregates were obtained after single cell inoculation on agarose microwells with efficient differentiation into the cardiac lineage using state-of-the-art protocols for directed differentiation via small molecules. With this soft lithography-based strategy, sufficient and reproducible numbers of stem cell-derived cardiomyocytes necessary for tissue engineering purposes can be realized in a highly controllable manner. Moreover, it might be useful for different cell types in any application that requires scalable and highly standardized aggregation.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

A9539
Agarose, BioReagent, for molecular biology, low EEO
A6013
Agarose, Type I, low EEO
A6877
Agarose, Medium EEO, for molecular biology
A0169
Agarose, Type I-A, low EEO
A4718
Agarose, High resolution, for molecular biology
A4679
Agarose, Low EEO, for Immunoelectrophoresis
A7431
Agarose, BioReagent, for molecular biology, Wide range/Standard 3:1
A9918
Agarose, Type II-A, Medium EEO
A0576
Agarose, Low EEO
A2790
Agarose, Wide range, for molecular biology
A9311
Agarose, Medium EEO, for electrophoresis
A2929
Agarose, For pulsed field electrophoresis running gel
A5030
Agarose, Ultra-low Gelling Temperature
A3643
Agarose, Type IV, Special High EEO
A2576
Agarose, Ultra-low Gelling Temperature, molecular biology grade
A9793
Agarose, High EEO
A7174
Agarose, High Gelling Temperature
A5304
Agarose, Special High EEO
A3054
Agarose, For pulsed field electrophoresis sample preparation
05066
Agarose, High EEO, for molecular biology
4B200
Sepharose® 4B, 45-165 μm bead diameter
6B100
Sepharose® 6B, 6% Beaded Agarose, 45-165 μm (wet), fractionation range 10,000-1,000,000 Da (dextrans)
U0507
Ultrogel® A4, aqueous ethanol suspension, 60-140 μm (wet bead diameter), exclusion limit 20,000,000 Da, fractionation range 55,000-9,000,000 Da (globular proteins)