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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and <t>cryopreservation.</t> (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.
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Image Search Results


Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and cryopreservation. (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.

Journal: Frontiers in Cellular Neuroscience

Article Title: Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics

doi: 10.3389/fncel.2024.1520992

Figure Lengend Snippet: Viability and OL lineage purity of cryopreserved OPCs. (A) Illustration of the pipeline for primary OL culture and cryopreservation. (B) Representative images of control and cryopreserved OPCs after 2 days of stabilization. Immunostaining for NG2 and Olig2. (C) Graph of viability measured by a dual-fluorescence cell counter utilizing AO/PI ( n = 4 biologically independent experiments per group, with three experimental groups. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). (D) Graph of OL lineage purity measured as Olig2 + DAPI + OPCs/DAPI + total cells (%) ( n = 4 biologically independent experiments per group, with three experimental groups. For each group, 10 images were analyzed, with approximately 75 DAPI + cells per image. Statistical analysis was performed using a two-sided Kruskal–Wallis test. Data are presented as mean ± SEM). OPC, oligodendrocyte progenitor cell; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; EGF, epidermal growth factor; T3, thyroid hormone T3; AO/PI, acridine orange/propidium iodide.

Article Snippet: The cell suspension in CELLBANKER 2 was transferred to cryopreservation vials (Thermo Fisher Scientific, #368632), two vials/flask, 1 mL/vial, and the vials were placed in a cell cryopreservation container (Thermo Fisher Scientific, #5100-0001) utilizing isopropanol.

Techniques: Control, Immunostaining, Fluorescence, Derivative Assay