Name: Isolamento e cultura de células Müller primárias da retina de ratos Sprague-Dawley (SD) (Vídeo) | JoVE Journal
Uploaded: 2025-06-17T14:20:00.000+00:00
Duration: 7 min 41 s
Description: Aprenda o isolamento e cultura de células Müller primárias da retina de ratos Sprague-Dawley. Este protocolo padronizado utiliza dissecação, digestão com tripsina e centrifugação para obter alta pureza celular, facilitando pesquisas em biologia retiniana e modelagem de doenças.

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<li>Bunt-Milam, A. H., Saari, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Immunocytochemical+localization+of+two+retinoid-binding+proteins+in+vertebrate+retina%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Immunocytochemical localization of two retinoid-binding proteins in vertebrate retina.</a> J Cell Biol. 97 (3), 703-712 (1983).</li>
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<li>Kolesnikov, A. V., Kiser, P. D., Palczewski, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Function+of+mammalian+M-cones+depends+on+the+level+of+CRALBP+in+M%C3%BCller+cells%5BTitle%5D)+AND+%22J+Gen+Physiol%22%5BJournal%5D)">Function of mammalian M-cones depends on the level of CRALBP in Müller cells.</a> J Gen Physiol. 153 (1), e202012675(2021).</li>
<li>Xue, Y., Shen, S. Q., Jui, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CRALBP+supports+the+mammalian+retinal+visual+cycle+and+cone+vision%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">CRALBP supports the mammalian retinal visual cycle and cone vision.</a> J Clin Invest. 125 (2), 727-738 (2015).</li>
<li>Pereiro, X., Beriain, S., Rodriguez, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Characteristics+of+whale+M%C3%BCller+glia+in+primary+and+immortalized+cultures%5BTitle%5D)+AND+%22Front+Neurosci%22%5BJournal%5D)">Characteristics of whale Müller glia in primary and immortalized cultures.</a> Front Neurosci. 16, 854278(2022).</li>
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<li>Zeng, Q., Xia, X. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Study+on+the+differentiation+of+retinal+ganglion+cells+from+rat+M%C3%BCller+cells+in+vitro%5BTitle%5D)+AND+%22Yan+Ke+Za+Zhi%22%5BJournal%5D)">Study on the differentiation of retinal ganglion cells from rat Müller cells in vitro.</a> Yan Ke Za Zhi. 46 (7), 615-620 (2010).</li>
<li>Backstrom, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Phenotypes+of+primary+retinal+macroglia%3A+Implications+for+purification+and+culture+conditions%5BTitle%5D)+AND+%22Exp+Eye+Res%22%5BJournal%5D)">Phenotypes of primary retinal macroglia: Implications for purification and culture conditions.</a> Exp Eye Res. 182, 85-92 (2019).</li>
<li>Liu, X., Tang, L., Liu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mouse+M%C3%BCller+cell+isolation+and+culture%5BTitle%5D)+AND+%22Bio+Protoc%22%5BJournal%5D)">Mouse Müller cell isolation and culture.</a> Bio Protoc. 7 (15), e2429(2017).</li>
<li>Pereiro, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Optimization+of+a+method+to+isolate+and+culture+adult+porcine%2C+rats+and+mice+M%C3%BCller+glia+in+order+to+study+retinal+diseases%5BTitle%5D)+AND+%22Front+Cell+Neurosci%22%5BJournal%5D)">Optimization of a method to isolate and culture adult porcine, rats and mice Müller glia in order to study retinal diseases.</a> Front Cell Neurosci. 14 (7), (2020).</li>
<li>Wang, M., Ma, W., Zhao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adaptive+M%C3%BCller+cell+responses+to+microglial+activation+mediate+neuroprotection+and+coordinate+inflammation+in+the+retina%5BTitle%5D)+AND+%22J+Neuroinflammation%22%5BJournal%5D)">Adaptive Müller cell responses to microglial activation mediate neuroprotection and coordinate inflammation in the retina.</a> J Neuroinflammation. 8, 173(2011).</li>
<li>Tomaszewski, R., Gad, M. S., Negoita, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+of+primary+mouse+retinal+glial+M%C3%BCller+cells%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Isolation of primary mouse retinal glial Müller cells.</a> J Vis Exp. (210), e66237(2024).</li>
<li>Li, Q., Cheng, Y., Zhang, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((TRPV4-induced+M%C3%BCller+cell+gliosis+and+TNF-%CE%B1+elevation-mediated+retinal+ganglion+cell+apoptosis+in+glaucomatous+rats+via+JAK2%2FSTAT3%2FNF-%CE%BAB+pathway%5BTitle%5D)+AND+%22J+Neuroinflammation%22%5BJournal%5D)">TRPV4-induced Müller cell gliosis and TNF-α elevation-mediated retinal ganglion cell apoptosis in glaucomatous rats via JAK2/STAT3/NF-κB pathway.</a> J Neuroinflammation. 18 (1), 271(2021).</li>
<li>McCarthy, K. D., de Vellis, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Preparation+of+separate+astroglial+and+oligodendroglial+cell+cultures+from+rat+cerebral+tissue%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue.</a> J Cell Biol. 85 (3), 890-902 (1980).</li>
<li>Reichenbach, A., Wolburg, H., Richter, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Membrane+ultrastructure+preservation+and+membrane+potentials+after+isolation+of+rabbit+retinal+glial+%28M%C3%BCller%29+cells+by+papain%5BTitle%5D)+AND+%22J+Neurosci+Methods%22%5BJournal%5D)">Membrane ultrastructure preservation and membrane potentials after isolation of rabbit retinal glial (Müller) cells by papain.</a> J Neurosci Methods. 32 (3), 227-233 (1990).</li>
<li>Harstad, H. K., Ringvold, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Scanning+and+transmission+electron+microscopy+of+M%C3%BCller+cells+isolated+from+rabbit+retina%5BTitle%5D)+AND+%22Graefes+Arch+Clin+Exp+Ophthalmol%22%5BJournal%5D)">Scanning and transmission electron microscopy of Müller cells isolated from rabbit retina.</a> Graefes Arch Clin Exp Ophthalmol. 223 (1), 29-34 (1985).</li>
<li>Guidry, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Isolation+and+characterization+of+porcine+M%C3%BCller+cells.+Myofibroblastic+dedifferentiation+in+culture%5BTitle%5D)+AND+%22Invest+Ophthalmol+Vis+Sci%22%5BJournal%5D)">Isolation and characterization of porcine Müller cells. Myofibroblastic dedifferentiation in culture.</a> Invest Ophthalmol Vis Sci. 37 (5), 740-752 (1996).</li>
<li>Qiu, A. W., Wang, N. Y., Yin, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Retinal+M%C3%BCller+cell-released+exosomal+miR-92a-3p+delivers+interleukin-17A+signal+by+targeting+Notch-1+to+promote+diabetic+retinopathy%5BTitle%5D)+AND+%22Invest+Ophthalmol+Vis+Sci%22%5BJournal%5D)">Retinal Müller cell-released exosomal miR-92a-3p delivers interleukin-17A signal by targeting Notch-1 to promote diabetic retinopathy.</a> Invest Ophthalmol Vis Sci. 66 (1), 1(2025).</li>
</ol>

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isolamento e cultura de células müller primárias da retina de ratos sprague-dawley (sd)

Isolation and Culture of Primary Retinal M&#252;ller Cells from Sprague-Dawley (SD) Rats

Depois que as células primárias são semeadas no frasco de cultura, as mudanças subsequentes no meio e a passagem ajudam a eliminar as RMCs pouco aderentes e a remover os detritos celulares gerados durante a passagem, enriquecendo assim a população de RMC na cultura. As CMRs foram identificadas com base em sua morfologia usando um microscópio óptico invertido (Figura 1) e coloração de hematoxilina e eosina (H&amp;E) (Figura 2). A ...

Isolamento e cultura de células Müller primárias da retina de ratos Sprague-Dawley (SD)

Retinal M&#252;ller cells (RMCs) play a crucial role in providing structural support and regulating various functions within the retina. As a core ...

isolation-culture-primary-retinal-mller-cells-from-sprague-dawley-sd

Research

JoVE Journal

Biology

Biologia

Neste vídeo, vamos demonstrar como isolar células-tronco na retina do epitélio ciliar do olho do rato e cultivá-las em cultura para dar forma clonal esferas da retina. As esferas que estão isolados possuem as propriedades cardeal de células-tronco: a auto-renovação e multipotencialidade.

Isolamento de Células-Tronco da retina do olho do rato

Milhões de pessoas sofrem de doenças degenerativas da retina que resultam em cegueira irreversível. Um elemento comum de muitas dessas doenças é a perda de células ganglionares da retina (RGCs). Este protocolo detalhado descreve o isolamento de RGC murinos primários por seleção positiva e negativa com citometria de fluxo.

Isolamento de células de gânglio retiniano murino primário (RGCs) por citometria de fluxo

O epithelium retinal do pigment (RPE) é um epitélio multi-funcional do olho. Aqui nós apresentamos um protocolo para estabelecer culturas de célula primária derivadas o RPE murino.

Culturas de células primárias do Mouse epithelium Retinal do Pigment

Este protocolo, que foi originalmente relatado por Fernandez-Godino et al. em 20161,descreve um método para isolar eficientemente e cultivar células RPE do mouse, que formam uma monocamada RPE funcional e polarizada dentro de uma semana em placas Transwell. O procedimento leva aproximadamente 3 horas.

Dissecção eficiente e cultura de células epiteliais de pigmento de retina do rato primário

Este manuscrito descreve um protocolo simplificado para o isolamento de células de epitélio pigmentado da retina (RPE) a partir de olhos de rato de forma stepwise. O protocolo inclui a enucleação e dissecação dos olhos do rato, seguido pelo isolamento, semeadura e cultivo de células RPE.

Isolamento das células epitélio pigmentadas do rato primário

Este protocolo descreve o procedimento para obtenção e cultivo de células epiteliais pigmentares primárias da retina (EPR) a partir de olhos suínos de origem local. Essas células servem como uma alternativa de alta qualidade às células-tronco e são adequadas para pesquisas in vitro da retina.

Isolamento de células epiteliais pigmentares primárias da retina suína para modelagem in vitro

Este manuscrito descreve um protocolo detalhado para isolar células gliais de Müller da retina de olhos de camundongos. O protocolo começa com a enucleação e dissecção dos olhos de camundongos, seguida de isolamento, semeadura e cultura de células de Müller.

Isolamento de células primárias de Müller glial da retina de camundongo

O epitélio pigmentar da retina (EPR) atua como uma barreira crucial entre a coróide e a retina, promovendo a saúde e a função dos tipos de células da retina, como os fotorreceptores. Aqui, descrevemos um protocolo simples e eficaz para isolar e cultivar EPR murino adulto.

Um método simplificado para isolamento e cultura de células epiteliais pigmentares da retina de camundongos adultos

A dissecção e crescimento de células de uma área do cérebro individuais facilita investigação de parâmetros celulares e fisiológicos. Nós descrevemos um método para cultivo de células primárias, que produz neurônios enriquecido culturas em um ambiente sem soro.

Cultura primária de neurônios do hipocampo de ratos recém-nascidos P0

Hepatócitos primários proporcionar uma ferramenta valiosa para avaliar as funções bioquímicas, moleculares, e metabólicas em um sistema fisiologicamente relevante experimental. Nós descrevemos um protocolo confiável para rato em perfusão de fígado in situ, que sempre gera hepatócitos viáveis ​​até to1.0 ×...

Isolamento e cultura primária de células hepáticas de ratos

Aprenda o isolamento e cultura de células Müller primárias da retina de ratos Sprague-Dawley. Este protocolo padronizado utiliza dissecação, digestão com tripsina e centrifugação para obter alta pureza celular, facilitando pesquisas em biologia retiniana e modelagem de doenças.

Isolamento e cultura de células Müller primárias da retina de ratos Sprague-Dawley (SD)

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