Journal ArticleDOI
A ROCK inhibitor permits survival of dissociated human embryonic stem cells
Kiichi Watanabe,Morio Ueno,Daisuke Kamiya,Ayaka Nishiyama,Michiru Matsumura,Takafumi Wataya,Jun Takahashi,Satomi Nishikawa,Shin-Ichi Nishikawa,Keiko Muguruma,Yoshiki Sasai +10 more
Reads0
Chats0
TLDR
Application of a selective Rho-associated kinase (ROCK) inhibitor, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency and facilitates subcloning after gene transfer, and enables SFEB-cultured hES Cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors.Abstract:
Poor survival of human embryonic stem (hES) cells after cell dissociation is an obstacle to research, hindering manipulations such as subcloning. Here we show that application of a selective Rho-associated kinase (ROCK) inhibitor1,2, Y-27632, to hES cells markedly diminishes dissociation-induced apoptosis, increases cloning efficiency (from ∼1% to ∼27%) and facilitates subcloning after gene transfer. Furthermore, dissociated hES cells treated with Y-27632 are protected from apoptosis even in serum-free suspension (SFEB) culture3 and form floating aggregates. We demonstrate that the protective ability of Y-27632 enables SFEB-cultured hES cells to survive and differentiate into Bf1+ cortical and basal telencephalic progenitors, as do SFEB-cultured mouse ES cells.read more
Citations
More filters
Journal ArticleDOI
Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.
Toshiro Sato,Robert G.J. Vries,Hugo J. Snippert,Marc van de Wetering,Nick Barker,Daniel E. Stange,Johan H. van Es,Arie Abo,Pekka Kujala,Peter J. Peters,Hans Clevers +10 more
TL;DR: It is concluded that intestinal cryptvillus units are self-organizing structures, which can be built from a single stem cell in the absence of a non-epithelial cellular niche.
Journal ArticleDOI
Cerebral organoids model human brain development and microcephaly
Madeline A. Lancaster,Magdalena Renner,Carol Anne Martin,Daniel Wenzel,Louise S. Bicknell,Matthew E. Hurles,Tessa Homfray,Josef M. Penninger,Andrew P. Jackson,Juergen A. Knoblich +9 more
TL;DR: A human pluripotent stem cell-derived three-dimensional organoid culture system that develops various discrete, although interdependent, brain regions that include a cerebral cortex containing progenitor populations that organize and produce mature cortical neuron subtypes is developed.
Journal ArticleDOI
Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling
Stuart M. Chambers,Christopher A. Fasano,Eirini P. Papapetrou,Mark J. Tomishima,Michel Sadelain,Lorenz Studer +5 more
TL;DR: Noggin/SB431542-based neural induction should facilitate the use of hES and hiPS cells in regenerative medicine and disease modeling and obviate the need for protocols based on stromal feeders or embryoid bodies.
Journal ArticleDOI
Reprogramming of human somatic cells to pluripotency with defined factors
In-Hyun Park,In-Hyun Park,Rui Zhao,Rui Zhao,Jason A. West,Jason A. West,Akiko Yabuuchi,Akiko Yabuuchi,Hongguang Huo,Hongguang Huo,Tan A. Ince,Paul H. Lerou,M. William Lensch,M. William Lensch,George Q. Daley,George Q. Daley +15 more
TL;DR: The data demonstrate that defined factors can reprogramme human cells to pluripotency, and establish a method whereby patient-specific cells might be established in culture.
Journal ArticleDOI
Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA
Luigi Warren,Philip D. Manos,Philip D. Manos,Tim Ahfeldt,Tim Ahfeldt,Yuin-Han Loh,Hu Li,Hu Li,Frank H. Lau,Wataru Ebina,Pankaj Mandal,Zachary D. Smith,Alexander Meissner,Alexander Meissner,George Q. Daley,Andrew S. Brack,James J. Collins,James J. Collins,James J. Collins,Chad A. Cowan,Thorsten M. Schlaeger,Thorsten M. Schlaeger,Derrick J. Rossi +22 more
TL;DR: It is shown that this approach can reprogram multiple human cell types to pluripotency with efficiencies that greatly surpass established protocols and represents a safe, efficient strategy for somatic cell reprogramming and directing cell fate that has broad applicability for basic research, disease modeling, and regenerative medicine.
References
More filters
Journal ArticleDOI
Embryonic Stem Cell Lines Derived from Human Blastocysts
James A. Thomson,Joseph Itskovitz-Eldor,Sander S. Shapiro,Michelle A. Waknitz,Swiergiel Jennifer J,Vivienne S. Marshall,Jeffrey M. Jones +6 more
TL;DR: Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages.
Journal ArticleDOI
Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro.
TL;DR: The derivation of pluripotent embryonic stem (ES) cells from human blastocysts is described, providing a model to study early human embryology, an investigational tool for discovery of novel growth factors and medicines, and a potential source of cells for use in transplantation therapy.
Journal ArticleDOI
Feeder-free growth of undifferentiated human embryonic stem cells.
Chunhui Xu,Margaret S. Inokuma,Jerrod Denham,Kathaleen Golds,Pratima Kundu,Joseph D. Gold,Melissa K. Carpenter +6 more
TL;DR: A successful feeder-free hES culture system in which undifferentiated cells can be maintained for at least 130 population doublings and are suitable for scaleup production is demonstrated.
Journal ArticleDOI
ROCKs: multifunctional kinases in cell behaviour
TL;DR: Recent experiments have defined new functions of ROCKs in cells, including centrosome positioning and cell-size regulation, which might contribute to various physiological and pathological states.
Journal ArticleDOI
Directed Differentiation of Embryonic Stem Cells into Motor Neurons
TL;DR: It is shown that developmentally relevant signaling factors can induce mouse embryonic stem cells to differentiate into spinal progenitor cells, and subsequently into motor neurons, through a pathway recapitulating that used in vivo.