RESUMO
The KCL016 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting splicing site of the VHL gene encoding von Hippel-Lindau tumor suppressor E3 ubiquitin protein ligase (676+3A>T). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Técnicas de Cultura de Células/métodos , Linhagem Celular/citologia , Células-Tronco Embrionárias Humanas/citologia , Mutação/genética , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Biomarcadores/metabolismo , Diferenciação Celular , Feminino , Humanos , Masculino , LinhagemRESUMO
The KCL021 human embryonic stem cell line was derived from an embryo donated for research that carried a ΔF508 mutation affecting the CFTR gene encoding the cystic fibrosis transmembrane conductance regulator. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , Células-Tronco Embrionárias Humanas/citologia , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Linhagem Celular , Hibridização Genômica Comparativa , Embrião de Mamíferos/citologia , Genótipo , Teste de Histocompatibilidade , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Mutação , LinhagemRESUMO
The KCL029 human embryonic stem cell line was derived from an embryo donated for research that carried a c.814T>C mutation in the WAS gene, which is linked to the Wiskott-Aldrich syndrome, a rare, inherited, X-linked, recessive disease characterized by immune dysregulation and microthrombocytopenia. The line is also carrier for a mutation p.N1152H in the gene encoding the cystic fibrosis transmembrane conductance regulator CFTR. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Células-Tronco Embrionárias Humanas/citologia , Proteína da Síndrome de Wiskott-Aldrich/genética , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Linhagem Celular , Hibridização Genômica Comparativa , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Embrião de Mamíferos/citologia , Genótipo , Teste de Histocompatibilidade , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Linhagem , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
The KCL024 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation in the NF1 gene encoding neurofibromin (c.3739-3742 ∆TTTG). Mutations in this gene have been linked to neurofibromatosis type 1, juvenile myelomonocytic leukemia and Watson syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Células-Tronco Embrionárias Humanas/citologia , Neurofibromina 1/genética , Diferenciação Celular , Células Cultivadas , Embrião de Mamíferos/citologia , Fertilização in vitro , Teste de Histocompatibilidade , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Microscopia de Fluorescência , Neurofibromatose 1/genética , Neurofibromatose 1/patologia , Linhagem , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição/metabolismoRESUMO
The KCL025 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation in the NF1 gene encoding neurofibromin (c.3739-3742 ΔTTTG). Mutations in this gene have been linked to neurofibromatosis type 1, juvenile myelomonocytic leukemia and Watson syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Células-Tronco Embrionárias Humanas/citologia , Neurofibromina 1/genética , Diferenciação Celular , Células Cultivadas , Embrião de Mamíferos/citologia , Fertilização in vitro , Teste de Histocompatibilidade , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Cariótipo , Masculino , Microscopia de Fluorescência , Linhagem , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
The KCL017 human embryonic stem cell line was derived from an embryo donated for research that carried an autosomal dominant mutation affecting splicing site of the VHL gene encoding von Hippel-Lindau tumor suppressor E3 ubiquitin protein ligase (676+3A>T). The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.
Assuntos
Células-Tronco Embrionárias Humanas/citologia , Ubiquitina-Proteína Ligases/genética , Diferenciação Celular , Células Cultivadas , Hibridização Genômica Comparativa , Embrião de Mamíferos/citologia , Fertilização in vitro , Genótipo , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Cariótipo , Masculino , Microscopia de Fluorescência , Linhagem , Polimorfismo Genético , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
AIM: Umbilical cord contains, within Wharton's jelly (WJ), multipotent mesenchymal stromal/stem cells (MSCs) of fetal origin that can be isolated and expanded in vitro with a minimal manipulation and very high efficiency. Our aim was to develop a highly reproducible protocol that has the unique potential to be scaled up and adapted to cGMP requirements for the use in cellular therapy. RESULTS: We found that derivation of WJ MSCs under defined conditions in low oxygen resulted in several folds higher populations of MSCA-1(+) cells (6.0-19.2%) when compared with WJ MSCs derived in the presence of serum (0.1-2.8%) or clinical-grade bone marrow (BM) MSCs cultured under atmospheric O2 (20%). We demonstrate that WJ MSCs derived following our protocol display antiproliferative activity similar to clinical-grade BM MSCs. We also show that these WJ MSCs can be differentiated into adipo-, chondro- and osteo-genic lineages. CONCLUSION: Easy accessibility, abundance and genetic 'naivety' make WJ MSCs logistically a more attractive source for clinical applications than BM MSCs.