Name: neuronal differentiation from mouse embryonic stem cells in vitro
Uploaded: 2020-06-02T14:00:00
Description: Watch this Scientific Journal Video about Neuronal Differentiation from Mouse Embryonic Stem Cells In vitro at JoVE.com

Dette arbejde blev støttet af National Natural Science Foundation of China (nr. 31501099) og midaldrende og unge i Uddannelsesafdelingen i Hubei-provinsen, Kina (nr. Q20191104). Og vi takker professor Wensheng Deng ved Wuhan University of Science and Technology for at levere musens embryonale stamcellelinjer A2lox.

1. Mus embryonale stamcellekultur
<ol><li>Forbered 0,1% gelatinebelagte cellekulturretter eller tallerkener.<ol>
<li>Der tilsættes 2 ml steriliseret 0,1% gelatine (0,1% m/v i vand) til 60 mm cellekulturretter. Rock forsigtigt for at sikre jævn belægning af cellekulturretterne.</li>
		<li>Sæt opvasken i en 5% CO2 inkubator ved 37 °C og lad belægningen i 1 time.</li>
		<li>Fjern 0,1% gelatineopløsningen, før du sår cellerne. BEMÆRK: Når gelatinen er fjernet, er det ikke nødvendigt at tørre elle...

<ol>
	<li>Vieira, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+stem+cell+differentiation+into+mature+neurons:+mechanisms+of+regulation+and+biotechnological+applications.">Neural stem cell differentiation into mature neurons: mechanisms of regulation and biotechnological applications.</a> Biotechnology Advances. 36 (7), 1946-1970 (2018).</li><li>Yang, J. R., Lin, Y. T., Liao, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+embryonic+stem+cells+on+Parkinson's+disease+therapy.">Application of embryonic stem cells on Parkinson's disease therapy.</a> Genomic Medicine, Biomarkers, and Health Sciences. 3 (1), 17-26 (2011).</li><li>Liu, C., Zhong, Y. W., Apostolou, A., Fang, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+differentiation+of+human+embryonic+stem+cells+as+an+in+vitro+tool+for+the+study+of+the+expression+patterns+of+the+neuronal+cytoskeleton+during+neurogenesis.">Neural differentiation of human embryonic stem cells as an in vitro tool for the study of the expression patterns of the neuronal cytoskeleton during neurogenesis.</a> Biochemical and Biophysical Research Communications. 439 (1), 154-159 (2013).</li><li>Khramtsova, E. A., Mezhevikina, L. M., Fesenko, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proliferation+and+differentiation+of+mouse+embryonic+stem+cells+modified+by+the+Neural+Growth+Factor+(NGF)+gene.">Proliferation and differentiation of mouse embryonic stem cells modified by the Neural Growth Factor (NGF) gene.</a> Biology Bulletin. 45 (3), 219-225 (2018).</li><li>Yoshimatsu, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluating+the+efficacy+of+small+molecules+for+neural+differentiation+of+common+marmoset+ESCs+and+iPSCs.">Evaluating the efficacy of small molecules for neural differentiation of common marmoset ESCs and iPSCs.</a> Neuroscience research. , (2019).</li><li>Kothapalli, C. R., Kamm, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+matrix+microenvironment+for+targeted+differentiation+of+embryonic+stem+cells+into+neural+and+glial+lineages.">3D matrix microenvironment for targeted differentiation of embryonic stem cells into neural and glial lineages.</a> Biomaterials. 34, 5995-6007 (2013).</li><li>Gazina, E. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+of+derivation+and+differentiation+of+mouse+embryonic+stem+cells+generating+synchronous+neuronal+networks.">Method of derivation and differentiation of mouse embryonic stem cells generating synchronous neuronal networks.</a> Journal of Neuroscience Methods. 293, 53-58 (2018).</li><li>Ohnuki, Y., Kurosawa, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+hanging+drop+culture+conditions+on+embryoid+body+formation+and+neuronal+cell+differentiation+using+mouse+embryonic+stem+cells:+Optimization+of+culture+conditions+for+the+formation+of+well-controlled+embryoid+bodies.">Effects of hanging drop culture conditions on embryoid body formation and neuronal cell differentiation using mouse embryonic stem cells: Optimization of culture conditions for the formation of well-controlled embryoid bodies.</a> Journal of Bioscience and Bioengineering. 115 (5), 571-574 (2013).</li><li>Wongpaiboonwattana, W., Stavridis, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+differentiation+of+mouse+embryonic+stem+cells+in+serum-free+monolayer+culture.">Neural differentiation of mouse embryonic stem cells in serum-free monolayer culture.</a> Journal of Visualized experiments. (99), e52823 (2015).</li><li>Li, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+optimized+method+for+neuronal+differentiation+of+embryonic+stem+cells+in+vitro.">An optimized method for neuronal differentiation of embryonic stem cells in vitro.</a> Journal of Neuroscience Methods. 330 (15), 108486 (2020).</li><li>Zhou, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+optimal+suspension+culture+time+for+the+osteoblastic+differentiation+of+human+induced+pluripotent+stem+cells+using+the+embryoid+body+method.">Investigation of the optimal suspension culture time for the osteoblastic differentiation of human induced pluripotent stem cells using the embryoid body method.</a> Biochemical and Biophysical Research Communications. 515 (4), 586-592 (2019).</li><li>Lu, J. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-trans+retinoic+acid+promotes+neural+lineage+entry+by+pluripotent+embryonic+stem+cells+via+multiple+pathways.">All-trans retinoic acid promotes neural lineage entry by pluripotent embryonic stem cells via multiple pathways.</a> BMC Cell Biology. 10, 57 (2009).</li><li>Kanungo, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic+acid+signaling+in+P19+stem+cell+differentiation.">Retinoic acid signaling in P19 stem cell differentiation.</a> Anticancer Agents in Medicinal Chemistry. 17 (9), 1184-1198 (2017).</li><li>Rochette-Egly, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic+acid+signaling+and+mouse+embryonic+stem+cell+differentiation:+Cross+talk+between+genomic+and+non-genomic+effects+of+RA.">Retinoic acid signaling and mouse embryonic stem cell differentiation: Cross talk between genomic and non-genomic effects of RA.</a> Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids. 1851 (1), 66-75 (2015).</li><li>Wang, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic+acid+maintains+self-renewal+of+murine+embryonic+stem+cells+via+a+feedback+mechanism.">Retinoic acid maintains self-renewal of murine embryonic stem cells via a feedback mechanism.</a> Differentiation. 76 (9), 931-945 (2008).</li><li>Wu, H. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic+acid-induced+upregulation+of+miR-219+promotes+the+differentiation+of+embryonic+stem+cells+into+neural+cells.">Retinoic acid-induced upregulation of miR-219 promotes the differentiation of embryonic stem cells into neural cells.</a> Cell Death Disease. 8, 2953 (2017).</li><li>Chen, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic+acid+regulates+germ+cell+differentiation+in+mouse+embryonic+stem+cells+through+a+Smad-dependent+pathway.">Retinoic acid regulates germ cell differentiation in mouse embryonic stem cells through a Smad-dependent pathway.</a> Biochemical and Biophysical Research Communications. 418 (3), 571-577 (2012).</li><li>Nakayama, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rapid+and+efficient+method+for+neuronal+induction+of+the+P19+embryonic+carcinoma+cell+line.">A rapid and efficient method for neuronal induction of the P19 embryonic carcinoma cell line.</a> Journal of Neuroscience Methods. 227, 100-106 (2014).</li><li>Bibel, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+mouse+embryonic+stem+cells+into+a+defined+neuronal+lineage.">Differentiation of mouse embryonic stem cells into a defined neuronal lineage.</a> Nature neuroscience. 7 (9), 1003-1009 (2004).</li><li>Coyle, D. E., Li, J., Baccei, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regional+Differentiation+of+Retinoic+Acid-Induced+Human+Pluripotent+Embryonic+Carcinoma+Stem+Cell+Neurons.">Regional Differentiation of Retinoic Acid-Induced Human Pluripotent Embryonic Carcinoma Stem Cell Neurons.</a> PLoS ONE. 6 (1), 16174 (2011).</li><li>Okada, Y., Shimazaki, T., Sobue, G., Okano, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinoic-acid-concentration-dependent+acquisition+of+neural+cell+identity+during+in+vitro+differentiation+of+mouse+embryonic+stem+cells.">Retinoic-acid-concentration-dependent acquisition of neural cell identity during in vitro differentiation of mouse embryonic stem cells.</a> Developmental Biology. 275, 124-142 (2004).</li></ol>

I denne undersøgelse etablerede vi en enkel og effektiv metode til neuronal differentiering fra mESC'er med lave omkostninger og let opnåede materialer. I denne metode kan 2 dages embryoid kropsdannelse efterfulgt af 6 dages RA induktion effektivt fremme differentieringen af mESC'er i NPC'ere (fase I-protokol 3). For fase II differentiering, N2B27 medium II (fase II-protokol 3) mest effektivt fremkalde differentiering fra NPC'ere i neuroner. For at sikre succes bør der lægges større vægt på flere kritiske trin.</p...

Embryonale stamceller (ESC'er) er pluripotente og kan differentiere sig til neurale prækursorceller (NPC'ere) og derefter til neuroner under visse betingelser1. ESC-baseret neurogenese giver den bedste platform til at efterligne neurogenese og tjener dermed som et nyttigt værktøj til udviklingsbiologiske undersøgelser og potentielt støtte i regenerativ medicin2,3. I de seneste årtier er mange strategier blevet rapporteret for at fremkalde embryonal neurogenese, såsom den transgene metode4, ved hjælp af små molekyler5, ved hjælp af en 3D-matrix mikromiljø6, og co-kultur teknikken7. De fleste af disse protokoller er dog enten begrænset eller svære at betjene, og de er derfor ikke egnede til brug i de fleste laboratorier.
For at finde en nem at betjene og billig metode til at opnå effektiv neural differentiering fra mESC'er blev der brugt en kombinatorisk screeningsstrategi her. Som beskrevet i figur 1blev hele processen med embryonal neurogenese opdelt i 2 faser. Fase I henviser til differentieringsprocessen fra MESC'er til NPC'ere, og fase II vedrører den efterfølgende differentiering fra NPC'er til neuroner. Baseret på principperne om nem betjening, lave omkostninger, let tilgængelige materialer og høj differentieringseffektivitet blev syv protokoller i fase I og tre protokoller i fase II valgt baseret på det traditionelle klæbende monolayerkultursystem eller embryoid kropsdannelsessystem8,9. Differentieringseffektiviteten af protokoller i begge faser blev evalueret ved hjælp af cellemorfologiobservation og immunfluorescensanalyse. Ved at kombinere den mest effektive protokol i hver fase etablerede vi den optimerede metode til neural differentiering fra MESC'er.

<table>
	<tbody>
		<tr>
			<td>Anti-Nestin antibody [Rat-401]</td>
			<td>Abcam</td>
			<td>Ab11306</td>
			<td>stored at -80 &#176;C, avoid repeated freezing and thawing</td>
		</tr>
		<tr>
			<td>Anti-&#946;-Tubulin III antibody produced in rabbit</td>
			<td>Sigma Aldrich</td>
			<td>T2200</td>
			<td>stored at -80 &#176;C, avoid repeated freezing and thawing</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488-Labeled Goat Anti-Mouse IgG</td>
			<td>Beyotime</td>
			<td>A0428</td>
			<td>stored at -20 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>B-27 Supplement (50X), serum free</td>
			<td>Gibco</td>
			<td>17504044</td>
			<td>stored at -20 &#176;C, and protect from light</td>
		</tr>
		<tr>
			<td>CHIR-99021 (CT99021)</td>
			<td>Selleck</td>
			<td>S1263</td>
			<td>stored at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Coverslips</td>
			<td>NEST</td>
			<td>801007</td>
			<td></td>
		</tr>
		<tr>
			<td>Cy3-Labeled Goat Anti-Rabbit IgG</td>
			<td>Beyotime</td>
			<td>A0516</td>
			<td>stored at -20 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>DME/F-12 1:1 (1x)</td>
			<td>HyClone</td>
			<td>SH30023.01B</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>HyClone</td>
			<td>SH30084.03</td>
			<td>stored at -20 &#176;C, avoid repeated freezing and thawing</td>
		</tr>
		<tr>
			<td>Fluorescence microscopy</td>
			<td>Olympus</td>
			<td>CKX53</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin</td>
			<td>Gibco</td>
			<td>CM0635B</td>
			<td>stored at room temperature</td>
		</tr>
		<tr>
			<td>GlutaMAX Supplement</td>
			<td>Gibco</td>
			<td>35050061</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Immunol Staining Primary Antibody dilution Buffer</td>
			<td>Beyotime</td>
			<td>P0103</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>KnockOut DMEM/F-12</td>
			<td>Gibco</td>
			<td>12660012</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>KnockOut Serum Replacement</td>
			<td>Gibco</td>
			<td>10828028</td>
			<td>stored at -20 &#176;C, avoid repeated freezing and thawing</td>
		</tr>
		<tr>
			<td>Leukemia Inhibitory Factor human</td>
			<td>Sigma</td>
			<td>L5283</td>
			<td>stored at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Mounting Medium With DAPI - Aqueous, Fluoroshield</td>
			<td>Abcam</td>
			<td>ab104139</td>
			<td>stored at 4 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>MEM Non-essential amino acids solution</td>
			<td>Gibco</td>
			<td>11140076</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>N-2 Supplement (100X)</td>
			<td>Gibco</td>
			<td>17502048</td>
			<td>stored at -20 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>Normal goat serum</td>
			<td>Jackson</td>
			<td>005-000-121</td>
			<td>stored at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Neurobasal Medium</td>
			<td>Gibco</td>
			<td>21103049</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Nonadhesive bacterial dish</td>
			<td>Corning</td>
			<td>3262</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (1X)</td>
			<td>HyClone</td>
			<td>SH30256.01B</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Penicillin/ Streptomycin Solution</td>
			<td>HyClone</td>
			<td>SV30010</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>PD0325901(Mirdametinib)</td>
			<td>Selleck</td>
			<td>S1036</td>
			<td>stored at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Retinoic acid</td>
			<td>Sigma</td>
			<td>R2625</td>
			<td>stored at -80 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>Strain 129 Mouse Embryonic Stem Cells</td>
			<td>Cyagen</td>
			<td>MUAES-01001</td>
			<td>Maintained in feeder-free culture system</td>
		</tr>
		<tr>
			<td>Stem-Cellbanker (DMSO free)</td>
			<td>ZENOAQ</td>
			<td>stem cellbanker DMSO free</td>
			<td>stored at -20 &#176;C, avoid repeated freezing and thawing</td>
		</tr>
		<tr>
			<td>Trypsin 0.25% (1X) Solution</td>
			<td>HyClone</td>
			<td>SH30042.01</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>T8787</td>
			<td></td>
		</tr>
		<tr>
			<td>2-Mercaptoethanol</td>
			<td>Gibco</td>
			<td>21985023</td>
			<td>stored at 4 &#176;C and protect from light</td>
		</tr>
		<tr>
			<td>4% paraformaldehyde</td>
			<td>Beyotime</td>
			<td>P0098</td>
			<td>stored at -20 &#176;C</td>
		</tr>
		<tr>
			<td>6 - well plate</td>
			<td>Corning</td>
			<td>3516</td>
			<td></td>
		</tr>
		<tr>
			<td>60 mm cell culture dish</td>
			<td>Corning</td>
			<td>430166</td>
			<td></td>
		</tr>
		<tr>
			<td>15 ml centrifuge tube</td>
			<td>NUNC</td>
			<td>339650</td>
			<td></td>
		</tr>
	</tbody>
</table>

neuronal differentiation from mouse embryonic stem cells in vitro

Den neurale differentiering af mus embryonale stamceller (MESCs) er et potentielt redskab til at belyse de vigtigste mekanismer, der er involveret i neurogenese og potentielt støtte i regenerativ medicin. Her etablerede vi en effektiv og billig metode til neuronal differentiering fra mESC'er in vitro, ved hjælp af strategien for kombinatorisk screening. Under de betingelser, der er defineret her, tillader den 2-dages embryoide kropsdannelse + 6-dages retinosyreinduktionsprotokol hurtig og effektiv differentiering fra mESC'er til neurale prækursorceller (NPC'er), som det ses ved dannelsen af godt stablet og neuritlignende A2lox og 129 derivater, der er Nestin-positive. Den sunde tilstand af embryoiøse kroppe og det tidspunkt, hvor retinosyre (RA) påføres, samt RA-koncentrationerne er kritiske i processen. I den efterfølgende differentiering fra NPC'ere til neuroner kunne N2B27 medium II (suppleret med Neurobasal medium) bedre understøtte langsigtet vedligeholdelse og modning af neuronale celler. Den præsenterede metode er meget effektiv, billig og nem at betjene, og kan være et kraftfuldt værktøj til neurobiologi og udviklingsbiologisk forskning.

2-dages embryoid kropsdannelse + 6-dages RA induktion fungerer bedst på at lede differentieringen af mESC'er i NPC'ere (fase I). For at bestemme den optimale protokol, der bedst fremmer differentieringen af MESC'er i NPC'ere (fase I), blev 7 protokoller testet på både A2lox og 129 mESC 'er (tabel 1), og differentieringsstatus for hver gruppe blev overvåget ved hjælp af lysmikroskop. Som det fremgår af figur 3A, viste de fleste A2lox- og 129-derivater under "2-dages emb...

Watch this Scientific Journal Video about Neuronal Differentiation from Mouse Embryonic Stem Cells In vitro at JoVE.com

Neuronal Differentiation from Mouse Embryonic Stem Cells In vitro

Her etablerede vi en billig og nem at betjene metode, der dirigerer hurtig og effektiv differentiering fra embryonale stamceller til neuroner. Denne metode er velegnet til popularisering blandt laboratorier og kan være et nyttigt værktøj til neurologisk forskning.

Neuronal differentiering fra mus embryonale stamceller In vitro

The neural differentiation of mouse embryonic stem cells (mESCs) is a potential tool for elucidating the key mechanisms involved in neurogenesis and ...

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Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors

Differentiering af musen embryonale stamceller i kortikale Interneuron prækursorer

GABAergic cortical interneurons are a heterogeneous population of cells that play critical roles in regulating the output of excitatory pyramidal neurons ...

Isolation and Culture of Embryonic Mouse Neural Stem Cells

Isolation og kultur af embryonale mus neurale stamceller

Neural stem cells (NSCs) are multipotent and can give rise to the three major cell types of the central nervous system (CNS). In vitro culture and ...

Purification of Prominin-1+ Stem Cells from Postnatal Mouse Cerebellum

Purification of Prominin-1+ Stem Cells from Postnatal Mouse Cerebellum

Rensning af Prominin-1+ Stamceller fra Postnatal Mouse Cerebellum

Most cerebellar neurons arise from two embryonic stem niches: a rhombic lip niche, which generates all the cerebellar excitatory glutamatergic neurons, ...

In Vitro Wedge Slice Preparation for Mimicking In Vivo Neuronal Circuit Connectivity

In Vitro Wedge Slice Forberedelse til efterligne In Vivo Neuronal Circuit Connectivity

In vitro slice electrophysiology techniques measure single-cell activity with precise electrical and temporal resolution. Brain slices must be relatively ...

In vitro Modeling for Neurological Diseases using Direct Conversion from Fibroblasts to Neuronal Progenitor Cells and Differentiation into Astrocytes

In vitro modellering for neurologiske sygdomme ved hjælp af direkte konvertering fra fibroblaster til neuronale stamceller og differentiering til astrocytter

Research on neurological disorders focuses primarily on the impact of neurons on disease mechanisms. Limited availability of animal models severely ...