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A Simple Method to Generate Super-sensitive AID (ssAID)-based Conditional Knockouts using CRISPR-based Gene Knockout in Various Vertebrate Cell Lines

Authors: Kohei Nishimura and Tatsuo Fukagawa, date: 07/20/2021, view: 44, Q&A: 0

[Abstract]

Inducing loss of function of a target protein using methods such as gene knockout is a powerful and useful strategy for analyzing protein function in cells. In recent years, the CRISPR/Cas-9-based gene knockout technology has been widely used across a variety of eukaryotes; however, this type of simple gene knockout strategy is not applicable to

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Generation of Mouse Pluripotent Stem Cell-derived Trunk-like Structures: An in vitro Model of Post-implantation Embryogenesis

Authors: Adriano Bolondi, Leah Haut, Seher Ipek Gassaloglu, Polly Burton, Helene Kretzmer, René Buschow, Alexander Meissner, Bernhard G. Herrmann and Jesse V. Veenvliet, date: 06/05/2021, view: 1244, Q&A: 0

[Abstract]

Post-implantation mammalian embryogenesis involves profound molecular, cellular, and morphogenetic changes. The study of these highly dynamic processes is complicated by the limited accessibility of in utero development. In recent years, several complementary in vitro systems comprising self-organized assemblies of mouse embryonic stem cells, such

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A Modified Semisolid Clonal Culture for Identification of B-1 and B-2 Progenitor Colony Forming Ability of Mouse Embryonic Hemogenic Endothelial Cells

Authors: Michihiro Kobayashi and Momoko Yoshimoto, date: 05/05/2020, view: 1618, Q&A: 0

[Abstract] The search for the origin of the first hematopoietic stem cells (HSCs) in the mouse embryo has been a hot topic in the field of developmental hematopoiesis. Detecting lymphoid potential is one of the supportive evidence to show the definitive hematopoietic activity of HSCs. However, the first B-lymphoid potential in the mouse embryos are reported ...

Differentiation of Mouse Embryonic Stem Cells to Neuronal Cells Using Hanging Droplets and Retinoic Acid

Authors: Jeroen Witteveldt and Sara Macias, date: 11/05/2019, view: 3214, Q&A: 0

[Abstract] Controlled differentiation of embryonic stem cells is an essential tool in stem cell research. In this protocol, we describe a simple differentiation protocol involving the induction of embryoid body formation in mouse embryonic stem cells (mESC) using hanging droplets, followed by differentiation into a neuronal lineage.

An Improved Staining Method for Low Signal LacZ Reporter Detection in Mouse Embryos

Authors: Xiaopeng Shen, Feng Xu, Shen Wu, Meng Li, Jingyi Zhang, Rui Liang and Yu Liu, date: 03/05/2019, view: 4105, Q&A: 0

[Abstract] In many fields of biology, especially in the field of developmental biology, LacZ reporter staining is an approach used to monitor gene expression patterns. In the LacZ reporter system, the LacZ gene is inserted in the endogenous location of the target gene via gene knock-in or by constructing a transgenic cassette in ...

Isolation of Neural Stem Cells from the Embryonic Mouse Hippocampus for in vitro Growth or Engraftment into a Host Tissue

Authors: Oksana Rybachuk, Olga Kopach, Tetyana Pivneva and Vitaliy Kyryk, date: 02/20/2019, view: 5388, Q&A: 0

[Abstract] For both stem cell research and treatment of the central nervous system disorders, neural stem/progenitor cells (NSPCs) represent an important breakthrough tool. In the expanded stem cell-based therapy use, NSPCs not only provide a powerful cell source for neural cell replacement but a useful model for developmental biology research. Despite ...

Quantification of Mouse Hematopoietic Progenitors’ Formation Using Time-lapse Microscopy and Image Analysis

Authors: Isabelle Bergiers, Christian Tischer, Özge Vargel Bölükbaşı and Christophe Lancrin, date: 01/05/2019, view: 3965, Q&A: 0

[Abstract] In vitro differentiation of mouse embryonic stem cells (mESCs) towards blood cells constitutes a well-established system to study the endothelial-to-hematopoietic transition (EHT) at the onset of blood development. Assessing the emergence of small non-adherent round blood cells in the culture without disturbing it is essential to evaluate ...

Small Molecule-Based Retinal Differentiation of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Authors: Jie Zhu and Deepak A. Lamba, date: 06/20/2018, view: 4931, Q&A: 0

[Abstract] Retinal degeneration leads to loss of light-sensing photoreceptors eventually resulting in vision impairment and impose a heavy burden on both patients and the society. Currently available treatment options are very limited and mainly palliative. Ever since the discovery of human pluripotent stem cell technologies, cell replacement therapy has ...

FACS-based Isolation of Neural and Glioma Stem Cell Populations from Fresh Human Tissues Utilizing EGF Ligand

Authors: Jessica Tome-Garcia, Fiona Doetsch and Nadejda M. Tsankova, date: 12/20/2017, view: 9569, Q&A: 0

[Abstract] Direct isolation of human neural and glioma stem cells from fresh tissues permits their biological study without prior culture and may capture novel aspects of their molecular phenotype in their native state. Recently, we demonstrated the ability to prospectively isolate stem cell populations from fresh human germinal matrix and glioblastoma ...

Transplantation of Embryonic Cortical Tissue into Lesioned Adult Brain in Mice

Authors: Cong Wang, Hao Gao and Shengxiang Zhang, date: 06/20/2017, view: 6234, Q&A: 0

[Abstract] Transplantation of embryonic cortical tissue for repairing the damaged brain has provided a potential therapy for brain injury and diseases. The grafted tissue can successfully survive and participate in reestablishing the functional neural circuit of the host brain. Transplantation surgery can be combined with fluorescently labeled transgenic ...

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