Cell Culture:Article Title: FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures.
Article Snippet: .. Materials VERO cells (ATCC CCL-81) Vero cell culture medium (see recipe) Zika virus strain, e.g., Uganda 1947 (MR 766, ATCC 1838) or Puerto Rico 2015 (PRVABC59, ATCC VR-1843) Vero cell infection medium (see recipe) 2× overlay medium (see recipe) 1% (w/v) DEAE dextran (MP Biomedicals, cat. no. 195133) in distilled water (sterilize by filtration) 5% (w/v) sodium bicarbonate (Fisher Scientific, cat. no. L-14639) in distilled water 2% Oxoid agar stock solution (see recipe) Janssens et al. 5 of 15 Current Protocols in Stem Cell Biology 19388969, 2019, 1, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpsc.65 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense 4% paraformaldehyde (PFA) in phosphate-buffered saline (Boston BioProducts, cat. no. BM-155) Dulbecco’s phosphate-buffered saline (DPBS, no calcium or magnesium, Invitrogen, cat. no. 14190144) 1% (w/v) crystal violet (Fisher Scientific, cat. no. C581-100) in distilled water Methanol 2D cerebral organoid culture of 7 days (see Basic Protocol 1) Cerebral organoid medium (see recipe) Penicillin/streptomycin (Sigma, cat. no. P4333) Fetal bovine serum (FBS, HyClone, cat. no. SH30071.03) EDTA solution (Invitrogen, cat. no. 15575-038) PermWash buffer (BD Biosciences, cat. no. 554723) Accutase (Innovative Cell Technologies, cat. no. AT104) DMEM/F12 (Gibco, cat. no. 11330-032) Fluorochrome-conjugated antibodies (see Table 1) T175 culture flasks 2-ml cryogenic vials 6-well flat-bottom cell culture plates Phase-contrast microscope 50-ml polypropylene centrifuge tubes Cell scrapers (optional) 1.5-ml microcentrifuge tubes Additional reagents and solutions for trypsinizing cells Prepare Zika virus supernatant 1.
Virus:Article Title: FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures.
Article Snippet: .. Materials VERO cells (ATCC CCL-81) Vero cell culture medium (see recipe) Zika virus strain, e.g., Uganda 1947 (MR 766, ATCC 1838) or Puerto Rico 2015 (PRVABC59, ATCC VR-1843) Vero cell infection medium (see recipe) 2× overlay medium (see recipe) 1% (w/v) DEAE dextran (MP Biomedicals, cat. no. 195133) in distilled water (sterilize by filtration) 5% (w/v) sodium bicarbonate (Fisher Scientific, cat. no. L-14639) in distilled water 2% Oxoid agar stock solution (see recipe) Janssens et al. 5 of 15 Current Protocols in Stem Cell Biology 19388969, 2019, 1, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpsc.65 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense 4% paraformaldehyde (PFA) in phosphate-buffered saline (Boston BioProducts, cat. no. BM-155) Dulbecco’s phosphate-buffered saline (DPBS, no calcium or magnesium, Invitrogen, cat. no. 14190144) 1% (w/v) crystal violet (Fisher Scientific, cat. no. C581-100) in distilled water Methanol 2D cerebral organoid culture of 7 days (see Basic Protocol 1) Cerebral organoid medium (see recipe) Penicillin/streptomycin (Sigma, cat. no. P4333) Fetal bovine serum (FBS, HyClone, cat. no. SH30071.03) EDTA solution (Invitrogen, cat. no. 15575-038) PermWash buffer (BD Biosciences, cat. no. 554723) Accutase (Innovative Cell Technologies, cat. no. AT104) DMEM/F12 (Gibco, cat. no. 11330-032) Fluorochrome-conjugated antibodies (see Table 1) T175 culture flasks 2-ml cryogenic vials 6-well flat-bottom cell culture plates Phase-contrast microscope 50-ml polypropylene centrifuge tubes Cell scrapers (optional) 1.5-ml microcentrifuge tubes Additional reagents and solutions for trypsinizing cells Prepare Zika virus supernatant 1.
Infection:Article Title: FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures.
Article Snippet: .. Materials VERO cells (ATCC CCL-81) Vero cell culture medium (see recipe) Zika virus strain, e.g., Uganda 1947 (MR 766, ATCC 1838) or Puerto Rico 2015 (PRVABC59, ATCC VR-1843) Vero cell infection medium (see recipe) 2× overlay medium (see recipe) 1% (w/v) DEAE dextran (MP Biomedicals, cat. no. 195133) in distilled water (sterilize by filtration) 5% (w/v) sodium bicarbonate (Fisher Scientific, cat. no. L-14639) in distilled water 2% Oxoid agar stock solution (see recipe) Janssens et al. 5 of 15 Current Protocols in Stem Cell Biology 19388969, 2019, 1, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpsc.65 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense 4% paraformaldehyde (PFA) in phosphate-buffered saline (Boston BioProducts, cat. no. BM-155) Dulbecco’s phosphate-buffered saline (DPBS, no calcium or magnesium, Invitrogen, cat. no. 14190144) 1% (w/v) crystal violet (Fisher Scientific, cat. no. C581-100) in distilled water Methanol 2D cerebral organoid culture of 7 days (see Basic Protocol 1) Cerebral organoid medium (see recipe) Penicillin/streptomycin (Sigma, cat. no. P4333) Fetal bovine serum (FBS, HyClone, cat. no. SH30071.03) EDTA solution (Invitrogen, cat. no. 15575-038) PermWash buffer (BD Biosciences, cat. no. 554723) Accutase (Innovative Cell Technologies, cat. no. AT104) DMEM/F12 (Gibco, cat. no. 11330-032) Fluorochrome-conjugated antibodies (see Table 1) T175 culture flasks 2-ml cryogenic vials 6-well flat-bottom cell culture plates Phase-contrast microscope 50-ml polypropylene centrifuge tubes Cell scrapers (optional) 1.5-ml microcentrifuge tubes Additional reagents and solutions for trypsinizing cells Prepare Zika virus supernatant 1.
Filtration:Article Title: FACS-Mediated Isolation of Neuronal Cell Populations From Virus-Infected Human Embryonic Stem Cell-Derived Cerebral Organoid Cultures.
Article Snippet: .. Materials VERO cells (ATCC CCL-81) Vero cell culture medium (see recipe) Zika virus strain, e.g., Uganda 1947 (MR 766, ATCC 1838) or Puerto Rico 2015 (PRVABC59, ATCC VR-1843) Vero cell infection medium (see recipe) 2× overlay medium (see recipe) 1% (w/v) DEAE dextran (MP Biomedicals, cat. no. 195133) in distilled water (sterilize by filtration) 5% (w/v) sodium bicarbonate (Fisher Scientific, cat. no. L-14639) in distilled water 2% Oxoid agar stock solution (see recipe) Janssens et al. 5 of 15 Current Protocols in Stem Cell Biology 19388969, 2019, 1, D ow nloaded from https://currentprotocols.onlinelibrary.w iley.com /doi/10.1002/cpsc.65 by U niversity O f T he Philipines D ilim an M ain L ibrary, W iley O nline L ibrary on [05/04/2024]. .. See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense 4% paraformaldehyde (PFA) in phosphate-buffered saline (Boston BioProducts, cat. no. BM-155) Dulbecco’s phosphate-buffered saline (DPBS, no calcium or magnesium, Invitrogen, cat. no. 14190144) 1% (w/v) crystal violet (Fisher Scientific, cat. no. C581-100) in distilled water Methanol 2D cerebral organoid culture of 7 days (see Basic Protocol 1) Cerebral organoid medium (see recipe) Penicillin/streptomycin (Sigma, cat. no. P4333) Fetal bovine serum (FBS, HyClone, cat. no. SH30071.03) EDTA solution (Invitrogen, cat. no. 15575-038) PermWash buffer (BD Biosciences, cat. no. 554723) Accutase (Innovative Cell Technologies, cat. no. AT104) DMEM/F12 (Gibco, cat. no. 11330-032) Fluorochrome-conjugated antibodies (see Table 1) T175 culture flasks 2-ml cryogenic vials 6-well flat-bottom cell culture plates Phase-contrast microscope 50-ml polypropylene centrifuge tubes Cell scrapers (optional) 1.5-ml microcentrifuge tubes Additional reagents and solutions for trypsinizing cells Prepare Zika virus supernatant 1.
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