Journal: Cell Proliferation

Article Title: Manufacturing clinical‐grade human induced pluripotent stem cell‐derived beta cells for diabetes treatment

doi: 10.1111/cpr.13232

Figure Lengend Snippet: Non‐exhaustive list of regulatory authorities and stem cell organizations involved in stem cell therapies

Article Snippet: Source companies with rights to reprogramming kits (non‐exhaustive list) , Thermo Fisher Scientific (CytoTune‐iPS Sendai Reprogramming Kit) , Thermo Fisher Scientific (Epi5 Episomal iPSC Reprogramming Kit) Lonza (Lonza L7 hiPSC Reprogramming and hPSC Culture System) Alstem (Episomal iPSC Reprogramming Kit) Creative Bioarray (QualiStem Episomal iPSC Reprogramming Kit) , Reprocell (Stemgent StemRNA 3rd Gen Reprogramming Kit) Creative Bioarray (QualiStem RNA iPSC Reprogramming Kit) Stem Cell Technologies (ReproRNA‐OKSGM) Merck (Simplicon RNA Reprogramming Kit).

Techniques:

Journal: PLoS ONE

Article Title: PI3K/AKT Signaling Pathway Is Essential for Survival of Induced Pluripotent Stem Cells

doi: 10.1371/journal.pone.0154770

Figure Lengend Snippet: Pictures of fibroblasts, fibroblast derived iPSC lines and iPSC-derived neurons were taken after treatment with 4 μM wortmannin at different time points (1 h, 2 h, 4 h, 6 h, 24 h) of fibroblasts, fibroblast deriveded iPSCs and iPSC-derived neurons. In contrast to both fibroblasts (NFH-46) (A) and iPSC-derived neurons (C), iPSCs (B) showed clear sensitivity to wortmannin induced apoptosis, resulting in a massive cell death with increasing time. The magnifications of all images are 40x.

Article Snippet: The iPSC line BIHi004-A was derived from normal human fibroblasts (NHDF-Ad-Der Fibroblasts, Lonza, Basel, Switzerland) using episomal vectors expressing Oct3/4, Sox2, Klf4, L-Myc, Lin28 mp53DD, and EBNA1 (Epi5 Episomal iPSC Reprogramming Kit, Life Technologies).

Techniques: Derivative Assay

Journal: PLoS ONE

Article Title: PI3K/AKT Signaling Pathway Is Essential for Survival of Induced Pluripotent Stem Cells

doi: 10.1371/journal.pone.0154770

Figure Lengend Snippet: (A) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in iPSCs treated for 24 h with 10 μM QVD-oph, 10ng/ml TRAIL, 4 μM wortmannin (Wort.), 4 μM L-779,450, 4 μM BMS, and 50 nM MLN-8237 (AKA-I). Insets: Histogram examples of cells treated with wortmannin or BMS as compared to controls (Con.). Sub-G1 cell populations are indicated (sG1). (B) Whole cell cycle analysis after treatment with small molecules mentioned in A with respect to the amount of different phases (sub-G1, G1, S, G2/M) in percent. (C) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in orginal fibroblasts treated for 24 h with 10 μM QVD-oph, 10ng/ml TRAIL, 4 μM wortmannin, 4 μM L-779,450, 4 μM BMS, and 10 μM AKA-I. Insets: Histogram examples of cells treated with wortmannin or BMS compared to controls. Sub-G1 cell populations are indicated (sG1). (D) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in iPSC-derived neurons treated with increasing concentrations (0.5, 1, 2, 4 μM) of wortmannin for 24 h. Insets: Histogram examples of cells treated with two different concentrations of wortmannin (0.5/2 μM) compared to controls. (E) Measurement of apoptosis after treatment with 4 μM wortmannin for 1 h compared to DMSO. (F) Apoptosis (percentage of sub-G1 cells) by 10 μM QVD-oph, 4 μM wortmannin or the combination of both in iPSCs after 24 h. (G) Percentages of different cell phases of iPSCs treated with 10 μM QVD-oph, 4 μM wortmannin or the combination of both for 24 h. Means and SDs are shown of three independent experiments in triplicates. Statistical significance (*; p < 0.05) is indicated for comparison of control cells and wortmannin-treated cells.

Article Snippet: The iPSC line BIHi004-A was derived from normal human fibroblasts (NHDF-Ad-Der Fibroblasts, Lonza, Basel, Switzerland) using episomal vectors expressing Oct3/4, Sox2, Klf4, L-Myc, Lin28 mp53DD, and EBNA1 (Epi5 Episomal iPSC Reprogramming Kit, Life Technologies).

Techniques: Cell Cycle Assay, Derivative Assay, Comparison, Control

Journal: PLoS ONE

Article Title: PI3K/AKT Signaling Pathway Is Essential for Survival of Induced Pluripotent Stem Cells

doi: 10.1371/journal.pone.0154770

Figure Lengend Snippet: (A,E) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in iPSCs treated for 24 h with 10ng/ml TRAIL, 4 μM L-779,450, 4 μM BMS, 50 nM MLN-8237 (AKA-I), 4 μM MK-2206, 4 μM wortmannin (Wort.). (B,F) Whole cell cycle analysis after treatment with small molecules mentioned in A with respect to the amount of different phases (sub-G1, G1, S, G2/M) in percent. (C,D) Histogram examples of cells treated with BMS, AKA-I or wortmannin as compared to controls (DMSO). Sub-G1 cell populations are indicated (sG1). (G) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in orginal fibroblasts (HFF and NHDF) treated for 24 h with 10ng/ml TRAIL, 4 μM L-779,450, 4 μM BMS, 50 nM MLN-8237 (AKA-I), 4 μM MK-2206, 4 μM wortmannin (Wort.). Insets: Histogram examples of cells treated with AKA-I or wortmannin compared to controls (DMSO). Sub-G1 cell populations are indicated (sG1). (H,I) Apoptosis (percentage of sub-G1 cells) was determined by cell cycle analysis in two iPSC-derived neurons treated for 24 h with 10ng/ml TRAIL, 4 μM L-779,450, 4 μM BMS, 50 nM MLN-8237 (AKA-I), 4 μM MK-2206, 4 μM wortmannin (Wort.). Insets: Histogram examples of cells treated with, BMS, AKA-I or wortmannin compared to controls (DMSO). Means and SDs are shown of three independent experiments in triplicates. Statistical significance (*; p < 0.05) is indicated for comparison of control cells and wortmannin-treated cells.

Article Snippet: The iPSC line BIHi004-A was derived from normal human fibroblasts (NHDF-Ad-Der Fibroblasts, Lonza, Basel, Switzerland) using episomal vectors expressing Oct3/4, Sox2, Klf4, L-Myc, Lin28 mp53DD, and EBNA1 (Epi5 Episomal iPSC Reprogramming Kit, Life Technologies).

Techniques: Cell Cycle Assay, Derivative Assay, Comparison, Control

Journal: PLoS ONE

Article Title: PI3K/AKT Signaling Pathway Is Essential for Survival of Induced Pluripotent Stem Cells

doi: 10.1371/journal.pone.0154770

Figure Lengend Snippet: Protein lysates of AD-iPSCs and iPSC-derived neurons (INC) were analyzed after treatment with 4μM wortmannin for different times. Fourty μg of protein each were separated by SDS-PAGE (12%). (A) Western blot analysis was used to monitor the expression of total and phosphorylated Akt (p-Akt, at serine 473). The anti-human LC3A recognizes both isoforms LC3I and LC3II and could also show phosphorylated LC3II. (B) iPSCs compared to iPSC-derived neurons and original fibroblasts. The anti-human Caspase-3 used recognizes only the cleaved active form of caspase-3. Beta-tubulin and Coomassie blue staining were used to confirm similar protein loading across samples.

Article Snippet: The iPSC line BIHi004-A was derived from normal human fibroblasts (NHDF-Ad-Der Fibroblasts, Lonza, Basel, Switzerland) using episomal vectors expressing Oct3/4, Sox2, Klf4, L-Myc, Lin28 mp53DD, and EBNA1 (Epi5 Episomal iPSC Reprogramming Kit, Life Technologies).

Techniques: Derivative Assay, SDS Page, Western Blot, Expressing, Staining