Derivation and Cloning of a Novel Rhesus Embryonic Stem Cell Line Stably Expressing Tau‐Green Fluorescent Protein

Embryonic stem cells (ESC) have the ability of indefinite self‐renewal and multilineage differentiation, and they carry great potential in cell‐based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC‐based transplantations in...

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Published inStem cells (Dayton, Ohio) Vol. 26; no. 6; pp. 1444 - 1453
Main Authors Wianny, Florence, Bernat, Agnieszka, Huissoud, Cyril, Marcy, Guillaume, Markossian, Suzy, Cortay, Véronique, Giroud, Pascale, Leviel, Vincent, Kennedy, Henry, Savatier, Pierre, Dehay, Colette
Format Journal Article
LanguageEnglish
Published Bristol John Wiley & Sons, Ltd 01.06.2008
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Abstract Embryonic stem cells (ESC) have the ability of indefinite self‐renewal and multilineage differentiation, and they carry great potential in cell‐based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC‐based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau‐Green fluorescent protein (tau‐GFP) is a powerful viable lineage tracer, allowing visualization of cell bodies, dendrites, and axons in exquisite detail. Here, we report the first rhesus monkey ESC line that ubiquitously and stably expresses tau‐GFP. First, we derived a new line of rhesus monkey ESC (LYON‐ES1) that show marker expression and cell cycle characteristics typical of primate ESCs. LYON‐ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ESC line stably expressing tau‐GFP that retains all the characteristics of the parental wild‐type line and is clonogenic. We show that neural precursors derived from the tau‐GFP ESC line are multipotent and that their fate can be precisely mapped in vivo after grafting in the adult rat brain. Disclosure of potential conflicts of interest is found at the end of this article.
AbstractList Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and carry great potential in cell based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety and efficacy of ESC based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau-green fluorescent protein (tau-GFP) is a powerful viable lineage tracer, allowing to visualize cell bodies, dendrites and axons in exquisite details. Here, we report the first rhesus monkey ESC line that ubiquituously and stably expresses tau-GFP. First, we derived a new line of rhesus monkey ESC (LYON-ES1), that show marker expression and cell cycle characteristics typical of primate ES cells. LYON-ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ES cell line stably expressing tau-GFP that retains all the characteristics of the parental wild type line and is clonogenic. We show that neural precursors derived from the tau-GFP ESC line are multipotent and that their fate can precisely be mapped in vivo after grafting in the adult rat brain.
Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and they carry great potential in cell-based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC-based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau-Green fluorescent protein (tau-GFP) is a powerful viable lineage tracer, allowing visualization of cell bodies, dendrites, and axons in exquisite detail. Here, we report the first rhesus monkey ESC line that ubiquitously and stably expresses tau-GFP. First, we derived a new line of rhesus monkey ESC (LYON-ES1) that show marker expression and cell cycle characteristics typical of primate ESCs. LYON-ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ESC line stably expressing tau-GFP that retains all the characteristics of the parental wild-type line and is clonogenic. We show that neural precursors derived from the tau-GFP ESC line are multipotent and that their fate can be precisely mapped in vivo after grafting in the adult rat brain. Disclosure of potential conflicts of interest is found at the end of this article.
Abstract Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and they carry great potential in cell-based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC-based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau-Green fluorescent protein (tau-GFP) is a powerful viable lineage tracer, allowing visualization of cell bodies, dendrites, and axons in exquisite detail. Here, we report the first rhesus monkey ESC line that ubiquitously and stably expresses tau-GFP. First, we derived a new line of rhesus monkey ESC (LYON-ES1) that show marker expression and cell cycle characteristics typical of primate ESCs. LYON-ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ESC line stably expressing tau-GFP that retains all the characteristics of the parental wild-type line and is clonogenic. We show that neural precursors derived from the tau-GFP ESC line are multipotent and that their fate can be precisely mapped in vivo after grafting in the adult rat brain. Disclosure of potential conflicts of interest is found at the end of this article.
Embryonic stem cells (ESC) have the ability of indefinite self‐renewal and multilineage differentiation, and they carry great potential in cell‐based therapies. The rhesus macaque is the most relevant preclinical model for assessing the benefit, safety, and efficacy of ESC‐based transplantations in the treatment of neurodegenerative diseases. In the case of neural cell grafting, tracing both the neurons and their axonal projections in vivo is essential for studying the integration of the grafted cells in the host brain. Tau‐Green fluorescent protein (tau‐GFP) is a powerful viable lineage tracer, allowing visualization of cell bodies, dendrites, and axons in exquisite detail. Here, we report the first rhesus monkey ESC line that ubiquitously and stably expresses tau‐GFP. First, we derived a new line of rhesus monkey ESC (LYON‐ES1) that show marker expression and cell cycle characteristics typical of primate ESCs. LYON‐ES1 cells are pluripotent, giving rise to derivatives of the three germ layers in vitro and in vivo through teratoma formation. They retain all their undifferentiated characteristics and a normal karyotype after prolonged culture. Using lentiviral infection, we then generated a monkey ESC line stably expressing tau‐GFP that retains all the characteristics of the parental wild‐type line and is clonogenic. We show that neural precursors derived from the tau‐GFP ESC line are multipotent and that their fate can be precisely mapped in vivo after grafting in the adult rat brain. Disclosure of potential conflicts of interest is found at the end of this article.
Author Markossian, Suzy
Wianny, Florence
Dehay, Colette
Bernat, Agnieszka
Kennedy, Henry
Leviel, Vincent
Marcy, Guillaume
Savatier, Pierre
Giroud, Pascale
Huissoud, Cyril
Cortay, Véronique
AuthorAffiliation 2 Service de Gynécologie et Obstétrique Hospices civils de Lyon Hôpital de la Croix Rousse 69004 Lyon,FR
3 Unité sous contrat plate-forme biotechnologique de cellules souches plate-forme PrimaStem INRA INSERM Université Claude Bernard - Lyon I 69500 Bron,FR
1 Institut cellule souche et cerveau INSERM : U846 Université Claude Bernard - Lyon I Centre de recherche Inserm 18, avenue du doyen lepine 69676 BRON CEDEX,FR
AuthorAffiliation_xml – name: 3 Unité sous contrat plate-forme biotechnologique de cellules souches plate-forme PrimaStem INRA INSERM Université Claude Bernard - Lyon I 69500 Bron,FR
– name: 1 Institut cellule souche et cerveau INSERM : U846 Université Claude Bernard - Lyon I Centre de recherche Inserm 18, avenue du doyen lepine 69676 BRON CEDEX,FR
– name: 2 Service de Gynécologie et Obstétrique Hospices civils de Lyon Hôpital de la Croix Rousse 69004 Lyon,FR
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  email: Colette.Dehay@inserm.fr
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Issue 6
Keywords Green Fluorescent Proteins
Cell Line
Teratoma
Genes
Blastocyst
Reporter
Macaca mulatta
Alkaline Phosphatase
Reverse Transcriptase Polymerase Chain Reaction
Lentivirus
tau Proteins
Stem Cell Transplantation
Animals
Zona Pellucida
Transfection
Cell Differentiation
Embryonic Stem Cells
Language English
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content type line 23
Agnieszka Bernat: data collection
Pascale Giroud: data collection
Cyril Huissoud: data collection
Colette Dehay: conception and design, data assembly and interpretation, financial support, manuscript writing
Guillaume Marcy: data collection and analysis
Suzy Markossian: provision of lentiviral vectors
Véronique Cortay: data collection
Pierre Savatier: conception and design, data interpretation, financial support, help with writing
Vincent Leviel: data collection
Florence Wianny: conception and design, data collection and interpretation, manuscript writing
Henry Kennedy : financial and administrative support, help with writing
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SSID ssj0014588
ssib005900054
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Snippet Embryonic stem cells (ESC) have the ability of indefinite self‐renewal and multilineage differentiation, and they carry great potential in cell‐based...
Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and they carry great potential in cell-based...
Abstract Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and they carry great potential in cell-based...
Embryonic stem cells (ESC) have the ability of indefinite self-renewal and multilineage differentiation, and carry great potential in cell based therapies. The...
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StartPage 1444
SubjectTerms Alkaline Phosphatase
Alkaline Phosphatase - metabolism
Animals
Blastocyst
Blastocyst - cytology
Cell Differentiation
Cell Line
Cellular Biology
Embryonic Stem Cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - enzymology
Embryonic Stem Cells - physiology
Embryonic Stem Cells - virology
Genes, Reporter
Green fluorescent protein
Green Fluorescent Proteins
Green Fluorescent Proteins - genetics
Lentivirus
Life Sciences
Macaca mulatta
Primates
Reverse Transcriptase Polymerase Chain Reaction
Rhesus monkey
Stem Cell Transplantation
Stem Cell Transplantation - methods
tau Proteins
tau Proteins - genetics
Teratoma
Teratoma - genetics
Teratoma - pathology
Transfection
Zona Pellucida
Zona Pellucida - physiology
Title Derivation and Cloning of a Novel Rhesus Embryonic Stem Cell Line Stably Expressing Tau‐Green Fluorescent Protein
URI https://onlinelibrary.wiley.com/doi/abs/10.1634%2Fstemcells.2007-0953
https://www.ncbi.nlm.nih.gov/pubmed/18356572
https://search.proquest.com/docview/19898583
https://inserm.hal.science/inserm-00409448
https://pubmed.ncbi.nlm.nih.gov/PMC2740840
Volume 26
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