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crx-gfp hesc line  (STEMCELL Technologies Inc)
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STEMCELL Technologies Inc crx-gfp hesc line
Generation of <t>hESC</t> derived retina organoids (CRX-GFP cell line) and RPE (H9 cell line). (A) Phase contrast images of organoid development up to day 56. (B–D) Progenitor gene expression analysis of CRX-GFP hESC-derived retina organoids ( n = 2) and undifferentiated stem cells ( n = 1). (B) OCT4 expression is absent in differentiated day 48 retina organoids. (C) Retina organoids (red) highly express genes indicative of primitive retina and photoreceptor progenitors. (D) Markers for mature, terminally differentiated retinal subpopulations. Retina organoids (red) contain markers for all the major retinal cell types, including photoreceptor progenitors, glia, ganglion cells, bipolar, horizontal, amacrine, and Müller cells. (E) Flow analysis shows that GFP expression is absent in the undifferentiated stem cells, early embryoid bodies, and young retina organoids. A distinct GFP+ population appears at day 70. Expression is highest day 78 and persists in the oldest analyzed day 91 organoids. (F) Examples of gates drawn used for FACS sorting of the GFP negative and GFP positive populations. (G) Post-FACS purities >90%. Cells were gated based on singlets, SSC vs. FSC profile, and if they were DAPI negative. Negative controls were undifferentiated CRX-GFP hESC (day 0). (H) Parylene only implant (no cells). (I) CPCB-RPE1 implant covered with hESC-RPE cells. (J) Enlargement of (I) to show details of hESC-RPE cells forming an epithelium (28–35 days culture).
Crx Gfp Hesc Line, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Generation of hESC derived retina organoids (CRX-GFP cell line) and RPE (H9 cell line). (A) Phase contrast images of organoid development up to day 56. (B–D) Progenitor gene expression analysis of CRX-GFP hESC-derived retina organoids ( n = 2) and undifferentiated stem cells ( n = 1). (B) OCT4 expression is absent in differentiated day 48 retina organoids. (C) Retina organoids (red) highly express genes indicative of primitive retina and photoreceptor progenitors. (D) Markers for mature, terminally differentiated retinal subpopulations. Retina organoids (red) contain markers for all the major retinal cell types, including photoreceptor progenitors, glia, ganglion cells, bipolar, horizontal, amacrine, and Müller cells. (E) Flow analysis shows that GFP expression is absent in the undifferentiated stem cells, early embryoid bodies, and young retina organoids. A distinct GFP+ population appears at day 70. Expression is highest day 78 and persists in the oldest analyzed day 91 organoids. (F) Examples of gates drawn used for FACS sorting of the GFP negative and GFP positive populations. (G) Post-FACS purities >90%. Cells were gated based on singlets, SSC vs. FSC profile, and if they were DAPI negative. Negative controls were undifferentiated CRX-GFP hESC (day 0). (H) Parylene only implant (no cells). (I) CPCB-RPE1 implant covered with hESC-RPE cells. (J) Enlargement of (I) to show details of hESC-RPE cells forming an epithelium (28–35 days culture).

Journal: Frontiers in Neuroscience

Article Title: Co-grafts of Human Embryonic Stem Cell Derived Retina Organoids and Retinal Pigment Epithelium for Retinal Reconstruction in Immunodeficient Retinal Degenerate Royal College of Surgeons Rats

doi: 10.3389/fnins.2021.752958

Figure Lengend Snippet: Generation of hESC derived retina organoids (CRX-GFP cell line) and RPE (H9 cell line). (A) Phase contrast images of organoid development up to day 56. (B–D) Progenitor gene expression analysis of CRX-GFP hESC-derived retina organoids ( n = 2) and undifferentiated stem cells ( n = 1). (B) OCT4 expression is absent in differentiated day 48 retina organoids. (C) Retina organoids (red) highly express genes indicative of primitive retina and photoreceptor progenitors. (D) Markers for mature, terminally differentiated retinal subpopulations. Retina organoids (red) contain markers for all the major retinal cell types, including photoreceptor progenitors, glia, ganglion cells, bipolar, horizontal, amacrine, and Müller cells. (E) Flow analysis shows that GFP expression is absent in the undifferentiated stem cells, early embryoid bodies, and young retina organoids. A distinct GFP+ population appears at day 70. Expression is highest day 78 and persists in the oldest analyzed day 91 organoids. (F) Examples of gates drawn used for FACS sorting of the GFP negative and GFP positive populations. (G) Post-FACS purities >90%. Cells were gated based on singlets, SSC vs. FSC profile, and if they were DAPI negative. Negative controls were undifferentiated CRX-GFP hESC (day 0). (H) Parylene only implant (no cells). (I) CPCB-RPE1 implant covered with hESC-RPE cells. (J) Enlargement of (I) to show details of hESC-RPE cells forming an epithelium (28–35 days culture).

Article Snippet: The first one, the cone-rod homeobox gene-green fluorescent protein (CRX-GFP) hESC line , was maintained with TeSR E8 media on Vitronectin XF (Stemcell Technologies) coated plates.

Techniques: Derivative Assay, Gene Expression, Expressing

Transplant histology at 2 months (A,B) and 6.5 months (C–E) post-surgery. (A,C) Co-graft area. (B,D) Host retina outside co-graft. (A–D) H&E staining. White asterisks indicate the center of rosettes (spherical arrangement of photoreceptors and other retinal layers around a central lumen). The transplant in (C) has grown around the parylene RPE implant (transplant #3 in , 197 dps). (E) Bestrophin (gold) and DAPI (blue) shows RPE and parylene in transplant (#2 in , 195 dps). Blue arrows point to parylene. Red arrows point to hESC-RPE. Yellow arrows point to host RPE. The separation of tissue is processing artifact. Scale bars = 100 μm (A–D) ; 20 μm (E) .

Journal: Frontiers in Neuroscience

Article Title: Co-grafts of Human Embryonic Stem Cell Derived Retina Organoids and Retinal Pigment Epithelium for Retinal Reconstruction in Immunodeficient Retinal Degenerate Royal College of Surgeons Rats

doi: 10.3389/fnins.2021.752958

Figure Lengend Snippet: Transplant histology at 2 months (A,B) and 6.5 months (C–E) post-surgery. (A,C) Co-graft area. (B,D) Host retina outside co-graft. (A–D) H&E staining. White asterisks indicate the center of rosettes (spherical arrangement of photoreceptors and other retinal layers around a central lumen). The transplant in (C) has grown around the parylene RPE implant (transplant #3 in , 197 dps). (E) Bestrophin (gold) and DAPI (blue) shows RPE and parylene in transplant (#2 in , 195 dps). Blue arrows point to parylene. Red arrows point to hESC-RPE. Yellow arrows point to host RPE. The separation of tissue is processing artifact. Scale bars = 100 μm (A–D) ; 20 μm (E) .

Article Snippet: The first one, the cone-rod homeobox gene-green fluorescent protein (CRX-GFP) hESC line , was maintained with TeSR E8 media on Vitronectin XF (Stemcell Technologies) coated plates.

Techniques: Staining