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Neural Crest Cells

Neural crest cells are a multipotent, migratory cell population that originates from the dorsal neural tube during embryonic development.
These cells give rise to a diverse array of cell types, including neurons, glia, melanocytes, craniofacial cartilage and bone, and endocrine cells.
Their remarkable plasticity and contributions to various tissues make neural crest cells a crucial area of study for developmental biologists and tissue engineers.
Explore the power of neural crest cells with PubCompare.ai's cutting-edge AI platform, which helps researchers locate the best experimental protocols from literature, preprints, and patents.
With intuitive comparisons, you can unlock the full potnetial of this dynamic cell type and take your research to new heights.

Most cited protocols related to «Neural Crest Cells»

Conditional Deletion of Wnt Signaling Regulator

The following transgenic mice used in this study were generated previously: EIIa-cre (Lakso et al., 1996 (link)), Wnt1-cre (Danielian et al., 1998 (link)), TOPGAL (DasGupta and Fuchs, 1999 (link)), and Pdx1-cre (Wells et al., 2007 (link)). The Wls floxed mice were crossed to the EIIa-cre, Wnt1-cre, and Pdx1-cre mice to generate conditional mice where Wls would be deleted in the germline, neural crest cells, or pancreatic progenitor cells, respectively. All mouse lines used in this study were genotyped by PCR using primers and protocols described previously. Yolk sacs from staged embryos or tail tips were digested overnight at 55°C in lysis buffer and genomic DNA extracted using a Kingfisher 96 Magnetic Particle Processor.

Carpenter A.C., Rao S., Wells J.M., Campbell K, & Lang R.A. (2010). Generation of Mice with a Conditional Null Allele for Wntless. Genesis (New York, N.Y. : 2000), 48(9), 554-558.

Publication 2010

Buffers Genome Germ Line Mice, Laboratory Mice, Transgenic Neural Crest Cells Oligonucleotide Primers Pancreas PDX1 protein, human Stem Cells Tail WNT1 protein, human Yolk Sac

Neural Crest Cell Migration and Proliferation

Digital images of the outgrowth cultures at 24–48 h were acquired by video microscopy. Then NIH Image software was used to measure the area of the outgrowth. This was obtained by taking the area of the entire explant and subtracting from it the area of the dense central neural tube mass. Since the neural tube fragment is attached to the substratum predominantly via its edges where neural crest cells are emerging, we controlled for differences in migration area resulting from random variations in the shape of the explant by dividing the outgrowth area (mm²) by the perimeter (mm) of the explant. This normalized number (in mm) is referred to as the migration index and provides an estimate of the migration rate of the neural crest cells. For monitoring cell proliferation, 48-h explant cultures were incubated with bromodeoxyuridine (BrdU) (10 μM/ml) for 2 h, fixed, and processed for immunodetection using an anti-BrdU antibody and a streptavidin peroxidase detection system (Calbiochem, San Diego, CA) used in conjunction with the True Blue™ peroxidase substrate (Kierkegaard and Perry Laboratories, Gaithersburg, MD). Labeled cells were counted with the aid of NIH Image. To determine the total cell number in the outgrowth after BrdU immunostaining, the cultures were further stained with hematoxylin. This permitted a total cell count to be obtained using NIH Image. The proliferation rate was then calculated as the percent of BrdU-labeled cells in the outgrowth.

Huang G.Y., Cooper E.S., Waldo K., Kirby M.L., Gilula N.B, & Lo C.W. (1998). Gap Junction–mediated Cell–Cell Communication Modulates Mouse Neural Crest Migration. The Journal of Cell Biology, 143(6), 1725-1734.

Publication 1998

5-bromouridine Antibodies, Anti-Idiotypic Cell Proliferation Cells Microscopy, Video Neural Crest Cells Perimetry Peroxidase Streptavidin true blue Tube, Neural

Directed Differentiation of hPSCs into Melanocytes

To derive neural crest cells and melanoblasts, hPSCs were plated at day −1 on matrigel-coated dishes in E8 medium and 10μM ROCKi. From day0 to day2 cells were cultured in E6 medium with 1ng/ml BMP4,10μM SB and 600nM CHIR. From day2 to day4, cells were cultured in E6 medium with 10μM SB + 1.5μM CHIR. From day 4 to day 6, cells were cultured in E6 medium with 1.5μM CHIR. From day 6 to day 11, cells were cultured in E6 medium with 1.5μM CHIR, 5ng/ml BMP4 and 100nM EDN3. Cells were FACS-sorted for cKIT and P75 expression, and double positive cells were further differentiated into melanocytes using melanocytes medium.
Detailed materials and methods are available in the supplementary materials.

Baggiolini A., Callahan S.J., Montal E., Weiss J.M., Trieu T., Tagore M.M., Tischfield S.E., Walsh R.M., Suresh S., Fan Y., Campbell N.R., Perlee S.C., Saurat N., Hunter M.V., Simon-Vermot T., Huang T.H., Ma Y., Hollmann T., Tickoo S.K., Taylor B.S., Khurana E., Koche R.P., Studer L, & White R.M. (2021). Developmental chromatin programs determine oncogenic competence in melanoma. Science (New York, N.Y.), 373(6559), eabc1048.

Publication 2021

Bone Morphogenetic Protein 4 Cells EDN3 protein, human Hyperostosis, Diffuse Idiopathic Skeletal matrigel Melanocyte Neural Crest Cells Proto-Oncogene Protein c-kit

Quantifying Neural Crest Cell Markers

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Publication 2010

Antibodies, Anti-Idiotypic Catenins Cell Nucleus Cells Clone Cells Cytoplasm DAPI Embryo Mesencephalon Migration, Cell Molecular Probes Neural Crest Neural Crest Cells Phalloidine SOX10 Transcription Factor Tube, Neural Vimentin

Single-cell RNA-seq of BMP-inhibited neural crest

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Publication 2021

Biopharmaceuticals cDNA Library Cells Chickens Cranium Division, Cell DNA, Complementary Embryo Flow Cytometry Genes Genome Love Neural Crest Cells Sequence Analysis

Most recents protocols related to «Neural Crest Cells»

Cultivation of Human Neuroblastoma Cell Lines

The human neuroblastoma SH-SY5Y and GI-ME-N cell lines were acquired from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, SK-N-AS and IMR-32 cell lines were obtained from the European Collection of Authenticated Cell Cultures (ECACC 94092302, 86041809, respectively) and the BE(2)-M17 cell line was acquired from the American Type Culture Collection (ATCC—Manassas, Virginia, United States). The SH-EP cell line was kindly provided by J. Roessler, Center for Pediatrics, Medical Center—University of Freiburg. Additional characteristics of the neuroblastoma cell lines used in this study are mentioned in Suppl. Table 1.
SH-SY5Y and BE(2)-M17 cells were maintained in 1:1 DMEM/F12 media (Gibco® Life Technologies) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco® Life Technologies), while GI-ME-N, SK-N-AS, SH-EP and IMR-32 cells were cultured in RPMI1640 media (Gibco® Life Technologies) supplemented with 10% FBS. Human embryonic stem cell-derived neural stem cells (hESC-NSCs) and neural crest cells (NCCs) were generated and maintained as previously described^{24 (link),59 (link),60 (link)}. No antibiotics were added to the culture media. All cell cultures were maintained at 37 °C in a 5% CO₂ humidified incubator.

Condurat A.L., Aminzadeh-Gohari S., Malnar M., Schider N., Opitz L., Thomas R., Menon V., Kofler B, & Pruszak J. (2023). Verteporfin-induced proteotoxicity impairs cell homeostasis and survival in neuroblastoma subtypes independent of YAP/TAZ expression. Scientific Reports, 13, 3760.

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Publication 2023

Antibiotics Cell Culture Techniques Cell Lines Cells Culture Media Europeans Homo sapiens Human Embryonic Stem Cells Neural Crest Cells Neural Stem Cells Neuroblastoma

hPSC Differentiation into SAPs

Utilizing a modified dual-SMAD inhibition differentiation protocol developed by the Studer laboratory at the Memorial Sloan Kettering Cancer Center, we performed in vitro differentiations of hPSCs into SAPs. Over the course of a 40-day differentiation, cells were cultured and sorted on day 16 for the CD49d maker (SOX10 positive cells), when cells are committed to trunc neural crest cells. Cells were harvested at the neural crest and hSAP stages. RNA was isolated from the collected cell pellets by lysing the cells in TRIzol Reagent (ThermoFisher catalog #15596018) and inducing phase separation with chloroform. Subsequently, RNA was precipitated with isopropanol and linear acrylamide and washed with 75% ethanol. The samples were resuspended in RNase-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 534–850 ng of total RNA with DV200% varying from 38–74% was used for ribosomal depletion and library preparation using the TruSeq Stranded Total RNA LT Kit (Illumina catalog #RS-122-1202) according to manufacturer’s instructions with 8 cycles of PCR. Samples were barcoded and run on a HiSeq 4000 in a 50 bp/50 bp paired end run, using the HiSeq 3000 / 4000 SBS Kit (Illumina). On average, 48 million paired reads were generated per sample and 35% of the data mapped to the transcriptome.

Decaesteker B., Louwagie A., Loontiens S., De Vloed F., Bekaert S.L., Roels J., Vanhauwaert S., De Brouwer S., Sanders E., Berezovskaya A., Denecker G., D’haene E., Van Haver S., Van Loocke W., Van Dorpe J., Creytens D., Van Roy N., Pieters T., Van Neste C., Fischer M., Van Vlierberghe P., Roberts S.S., Schulte J., Ek S., Versteeg R., Koster J., van Nes J., Zimmerman M., De Preter K, & Speleman F. (2023). SOX11 regulates SWI/SNF complex components as member of the adrenergic neuroblastoma core regulatory circuitry. Nature Communications, 14, 1267.

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Publication 2023

Acrylamide cDNA Library Chloroform Endoribonucleases Ethanol Isopropyl Alcohol Malignant Neoplasms Neural Crest Neural Crest Cells Pellets, Drug Psychological Inhibition Ribosomes SKAP2 protein, human SOX10 Transcription Factor Torso Transcriptome trizol

Conditional overexpression of Dlx2 in neural crest cells

We obtained wnt1^cre mice from the Jackson laboratory. The Rosa26^{CAG-LSL-Dlx2−3xFlag} mouse was constructed by the Shanghai Model Organisms Center, Inc. (Shanghai, China). To generate wnt1^cre; Rosa26^Dlx2/- mice, which could specifically overexpress Dlx2 in neural crest cells, we mated wnt1^cre mice with Rosa26^{CAG-LSL-Dlx2−3xFlag} mice. Wildtype C57BL/6J mice were purchased from Shanghai Jihui Laboratory Animal Care Co. Ltd. (Shanghai, China). All mice were maintained under SPF conditions at the Animal Center of the Ninth People’s Hospital affiliated with Shanghai Jiao Tong University School of Medicine. The day of the appearance of a vaginal plug was defined as E0.5 in all timed pregnancies. Embryos at the E10.5 and E12.5 stages (12:00 h of the day when the vaginal plug was detected was counted as E0.5) and P0 pups were collected for subsequent experiments. All animal experiments were approved by the Animal Care and Usage Committee of the Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of medicine.

Sun J., Zhang J., Bian Q, & Wang X. (2023). Effects of Dlx2 overexpression on the genes associated with the maxillary process in the early mouse embryo. Frontiers in Genetics, 14, 1085263.

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Publication 2023

Animal Diseases Animals Animals, Laboratory Embryo Mice, House Mice, Inbred C57BL Mice, Laboratory Neural Crest Cells Pregnancy Vagina

Induced Mesenchymal Stem Cell Generation

iMSCs were generated by inducing hiPSCs into neural crest cells (NCCs), and then into iMSCs (Kamiya et al., 2022 (link)), which were then maintained in a plastic dish coated with Fibronectin (Chemicon, FC010-10 MG) and fed with PRIME-XV^®MSC Expansion XSFM (IrvineScientific, 91149). Analyses were performed during passages 3—5.

Al-Akashi Z., Zujur D., Kamiya D., Kato T J.r., Kondo T, & Ikeya M. (2023). Selective vulnerability of human-induced pluripotent stem cells to dihydroorotate dehydrogenase inhibition during mesenchymal stem/stromal cell purification. Frontiers in Cell and Developmental Biology, 11, 1089945.

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Publication 2023

FN1 protein, human Human Induced Pluripotent Stem Cells Hyperostosis, Diffuse Idiopathic Skeletal Neural Crest Cells

Comprehensive scMST and Bulk RNAseq Protocols for Studying Midbrain Neural Crest

For scMST, samples were collected on Whatman filter papers, fixed in 4% PFA overnight, and washed 3 times with PBS-0.2% triton, dehydrated in ethanol and stored in −80°C. The embryos we incubated through a sucrose gradient (5% 30 min, 15% 4h at 4°C and embedded into OCT as previously described ^{6 (link),7 (link)} and 20 μm thick transverse cryosections were cut from the midbrain level onto silane coated coverslips.
For bulk RNAseq experiments, embryos were collected on Whatman filter papers in PBS and midbrain level neural plate border and specified neural crest cells, respectively, from left and right sides were manually microdissected using tungsten needles, immediately placed into RNA Lysis Buffer (RNAqueous^™-4PCR Total RNA Isolation Kit, Thermofisher, Waltham, MA, USA) and stored at −80 °C. Replicates consisted of pooled tissue from at least five to seven embryos. Four replicates were collected per stage. For HH5 and HH6 replicates, rectangular patches lateral to the primitive streak were collected according to midbrain neural crest fate mapping as described ^{13 (link)}. For premigratory stages beyond HH6, neural plate borders or neural fold apices corresponding to the midbrain neural crest domain were excised. Beyond 7ss, dorsal neural tubes containing neural crest from the midline to migratory front were collected.
The scRNAseq samples were collected by dissecting midbrain slices (covering all germ layers) from the desired stage by using micro scissors, which were pooled from three to four embryos per sample and dissociated with a multi tissue dissociation kit (kit 3,130-110-204, Miltenyi Biotec,) at 37°C for 20 min with intermittent pipetting to achieve a single cell suspension.

Pajanoja C., Hsin J., Olinger B., Schiffmacher A., Abrams S., Dapkunas A., Zainul Z., Doyle A.D., Martin D, & Kerosuo L. (2023). Maintenance of pluripotency in the entire ectoderm enables neural crest formation. Research Square.

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Publication Preprint 2023

Buffers Cells Cryoultramicrotomy Dietary Fiber Embryo Ethanol Germ Layers isolation Mesencephalon Needles Neural Crest Neural Crest Cells Neural Fold Neural Plate Primitive Streak Silanes Single-Cell RNA-Seq Sucrose Tissues Tube, Neural Tungsten

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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.

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What are the key characteristics of neural crest cells?

Neural crest cells are a unique and versatile cell population that originate from the dorsal neural tube during embryonic development. These cells are multipotent, meaning they can give rise to a diverse array of cell types, including neurons, glia, melanocytes, craniofacial cartilage and bone, and endocrine cells. Their remarkable plasticity and contributions to various tissues make neural crest cells a crucial area of study for developmental biologists and tissue engineers.

How can neural crest cells be utilized in research and applications?

Neural crest cells have a wide range of applications in research and medicine. Their ability to differentiate into various cell types makes them valuable for regenerative medicine, tissue engineering, and the study of developmental processes. Researchers can harness the potential of neural crest cells to generate replacement cells for damaged or diseased tissues, as well as to better understand the mechanisms underlying congenital disorders and cancers that may involve these cells.

What are some common challenges in working with neural crest cells?

One of the main challenges in working with neural crest cells is the complexity of their developmental pathways and the need to carefully control the differentiation process. Ensuring the purity and homogeneity of neural crest cell populations can also be a significant hurdle, as these cells can easily interconvert with other cell types. Additionally, the precise culture conditions and signaling cues required to maintain the desired phenotypes of neural crest cells can vary depending on the specific application.

How can PubCompare.ai assist researchers working with neural crest cells?

PubCompare.ai's AI-driven platform can be an invaluable tool for researchers working with neural crest cells. The platfomr allows you to more efficiently screen the literature on neural crest cell protocols, leveraging AI to pinpoint critical insights that can help you identify the most effective protocols for your research goals. By comparing the effectiveness of different protocols, the platform's analysis can highlight key differences and enable you to choose the best option for reproducibility and accuracy, ultimately enhancing your neural crest cell research.

Are there different types or variations of neural crest cells?

Yes, there are several variations and subpopulations of neural crest cells that have been identified. For example, trunk neural crest cells and cephalic neural crest cells have distinct developmental potentials and contribute to different tissues. Additionally, neural crest cells can be further classified based on their migratory pathways, temporal emergence, and molecular signatures. Understanding these nuanced differences is important for researchers to tailor their experimental approaches and applications to the specific neural crest cell subtype they are studying.

More about "Neural Crest Cells"

Neural crest cells (NCCs) are a remarkable population of multipotent, migratory cells that originate from the dorsal neural tube during embryonic development.
These dynamic cells have the remarkable ability to differentiate into a diverse array of cell types, including neurons, glial cells, melanocytes, craniofacial cartilage and bone, and endocrine cells.
Their remarkable plasticity and wide-ranging contributions to various tissues make NCCs a crucial area of study for developmental biologists and tissue engineers.
Researchers can explore the power of NCCs using PubCompare.ai's cutting-edge AI platform.
This innovative tool helps scientists locate the best experimental protocols from the literature, preprints, and patents, enhancing the reproducibility and accuracy of NCC-related studies.
With intuitive comparisons, researchers can unlock the full potential of this dynamic cell type and take their work to new heights.
To support NCC research, scientists often utilize specialized cell culture media, such as Fetal Bovine Serum (FBS), Dulbecco's Modified Eagle Medium (DMEM), and DMEM/F12.
Additionally, tools like the RNeasy Mini Kit can facilitate efficient RNA extraction, while supplements like B27 can promote the growth and differentiation of NCC-derived cell types.
Techniques such as flow cytometry using the FACSAria III system can help researchers isolate and analyze NCC populations.
Other key resources for NCC research include growth factors like Basic Fibroblast Growth Factor (bFGF) and the SMART-Seq v4 Ultra Low Input cDNA Kit for high-quality transcriptomic analysis.
The Accumax cell dissociation reagent can also be employed to gently dissociate NCC-containing tissues.
By leveraging these specialized tools and techniques, researchers can delve deeper into the fascinating world of neural crest cells and unlock new insights that can drive advancements in developmental biology and regenerative medicine.
OtherTerms: Neural crest, multipotent cells, migratory cells, dorsal neural tube, cell types, neurons, glia, melanocytes, craniofacial, cartilage, bone, endocrine, developmental biology, tissue engineering, PubCompare.ai, experimental protocols, literature, preprints, patents, cell culture, FBS, DMEM, DMEM/F12, RNeasy Mini Kit, B27 supplement, Accumax, FACSAria III, bFGF, SMART-Seq v4 Ultra Low Input cDNA Kit, AF1494