Directed Differentiation of Pluripotent Stem Cells

hESC lines H1 and H9 were obtained from WiCell Research Institute. iPS-MSC^{25 (link)} (derived from human mesenchymal stem cells) and iPS-Fib2^{25 (link)} (derived from human dermal fibroblasts) were a kind gift from George Q. Daley at Children’s Hospital Boston. Essential 8 medium (E8)^{5 (link)}, 0.5 mM EDTA, Accutase, ProLong® Antifade reagents, LIVE/DEAD® Cell Viability staining kit, Click-iT® EdU Alexa Fluor® 594 Imaging Kit and Alkaline Phosphatase Live Stain kit were obtained from Life Technologies. Small molecules Y-27632 (ROCK inhibitor, or RI), SB431542, and LDN193189 were from Selleckchem. Matrigel was obtained from BD Biosciences. PNIPAAm-PEG polymer (Mebiol Gel) was from Cosmo Bio, USA. SCID Beige mice were from Charles River Laboratory. Finally, the following antibodies and dilutions were used: Oct4 and Nanog (Santa Cruz Biotech, 1:100); FOXA2 or HNF3β (Santa Cruz Biotech, 1:200); Nestin (Millipore, 1:200); αSMA (Abcom, 1:200).

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Lei Y., Jeong D., Xiao J, & Schaffer D.V. (2014). Developing Defined and Scalable 3D Culture Systems for Culturing Human Pluripotent Stem Cells at High Densities. Cellular and molecular bioengineering, 7(2), 172-183.

Publication 2014

hESC lines H1 and H9 Accutase ProLong® Antifade reagents Click-iT® EdU Alexa Fluor® 594 Imaging Kit Alkaline Phosphatase Live Stain kit Y-27632 SB431542 LDN193189 Matrigel SCID Beige mice Nanog FOXA2 HNF3β Nestin

Accutase Alexa 594 Alkaline phosphatase Antibodies Cell viability Dilutions Edta Fibroblasts Hesc Human Ldn193189 Matrigel Mesenchymal stem cells Nestin Oct4 Pnipaam Polymer River Sb431542 Scid mice Stain Y 27632 αsma

Corresponding Organization : QB3

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Variable analysis

independent variables

HESC lines H1 and H9
IPS-MSC (derived from human mesenchymal stem cells)
IPS-Fib2 (derived from human dermal fibroblasts)

dependent variables

Cell viability
Proliferation

control variables

Essential 8 medium (E8)
0.5 mM EDTA
Accutase
ProLong® Antifade reagents
LIVE/DEAD® Cell Viability staining kit
Click-iT® EdU Alexa Fluor® 594 Imaging Kit
Alkaline Phosphatase Live Stain kit
Y-27632 (ROCK inhibitor, or RI)
SB431542
LDN193189
Matrigel
PNIPAAm-PEG polymer (Mebiol Gel)
SCID Beige mice
Antibodies: Oct4, Nanog, FOXA2 or HNF3β, Nestin, αSMA

Annotations

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