Developmental Biology

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    Protocols in Current Issue
    Application of a Modified Smart-seq2 Sample Preparation Protocol for Rare Cell Full-length Single-cell mRNA Sequencing to Mouse Oocytes
    Authors:  Rebecca S. Treger, Scott D. Pope, Xiaojun Xing and Akiko Iwasaki, date: 08/20/2019, view: 258, Q&A: 0
    [Abstract] Endogenous retroviruses (ERV) are transposable retroelements that form ~10% of the murine genome and whose family members are differentially expressed throughout embryogenesis. However, precise regulation of ERV in germ cells remains unclear. To investigate ERV expression in oocytes, we adapted a single-cell mRNA-sequencing library preparation ...
    CRISPR/Cas9 + AAV-mediated Intra-embryonic Gene Knocking in Mice
    [Abstract] Intra-embryo genome editing by CRISPR/Cas9 has enabled rapid generation of gene knockout animals. However, large fragment knock-in directly into embryos’ genome is still difficult, especially without microinjection of donor DNA. Viral vectors are good transporters of knock-in donor DNA for cell lines, but seemed unsuitable for pre-implantation ...
    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: 1101, 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 ...
    Heterochronic Phenotype Analysis of Hypodermal Seam Cells in Caenorhabditis elegans
    Authors:  Yon Ju Ji and Jiou Wang, date: 01/05/2019, view: 1514, Q&A: 0
    [Abstract] Heterochrony refers to changes in the timing of developmental events, and it is precisely regulated in the organisms by the heterochronic genes such as C. elegans lin-4 and let-7. Mutations in these genes cause precocious or retarded development of certain cell lineages. With well-defined cell lineages, C. elegans ...
    Quantification of Mouse Hematopoietic Progenitors’ Formation Using Time-lapse Microscopy and Image Analysis
    [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 ...
    Notochord Injury Assays that Stimulate Transcriptional Responses in Zebrafish Larvae
    [Abstract] Zebrafish have become an increasingly important model organism in the field of wound healing and regenerative medicine, due to their high regenerative capacity coupled with high-resolution imaging in living animals. In a recent study, we described multiple physical and chemical methods to induce notochord injury that led to highly specific ...
    Generation of BMEC Lines and in vitro BMEC-HSPC Co-culture Assays
    Authors:  Peipei Guo and Shahin Rafii, date: 11/05/2018, view: 1500, Q&A: 0
    [Abstract] Endothelial cells (ECs) sustain the self-renewal and regeneration of adult hematopoietic stem and progenitor cells (HSPCs) via deployment of EC-derived paracrine factors, termed as angiocrine factors. Generation of durable ex vivo vascular niche that maintains EC identity and preserves the angiocrine profile of organ of origin offers ...
    Detection of Cell Death in Planarians
    Authors:  Nídia de Sousa and Teresa Adell, date: 10/05/2018, view: 1834, Q&A: 0
    [Abstract] Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the ...
    Human Endothelial Cell Spheroid-based Sprouting Angiogenesis Assay in Collagen
    Authors:  Fabian Tetzlaff and Andreas Fischer, date: 09/05/2018, view: 4242, Q&A: 0
    [Abstract] Angiogenesis, the formation of new blood vessels from pre-existing ones plays an important role during organ development, regeneration and tumor progression. The spheroid-based sprouting assay is a well-established and robust method to study the influence of genetic alterations or pharmacological compounds on capillary-like tube formation of ...
    Pollen Germination and Pollen Tube Growth of Arabidopsis thaliana: In vitro and Semi in vivo Methods
    Authors:  Hugh Dickinson, Josefina Rodriguez-Enriquez and Robert Grant-Downton , date: 08/20/2018, view: 4449, Q&A: 2
    [Abstract] Studies of pollen germination and post-germination development are not only essential for understanding plant reproduction but also are an excellent model system for tip-based growth. Here we describe easy, reproducible methods for germination and growth of pollen from the model plant Arabidopsis thaliana in artificial conditions. Our ...



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