Journal: Stem Cell Reports

Article Title: Polysaccharides from Ganoderma lucidum Promote Cognitive Function and Neural Progenitor Proliferation in Mouse Model of Alzheimer's Disease

doi: 10.1016/j.stemcr.2016.12.007

Figure Lengend Snippet: G. lucidum Polysaccharides Increase NPC Proliferation and Self-Renewal In Vitro (A and B) Monolayer adult hippocampal NPC cultures were treated with GLP of different concentrations for 24 hr in culture medium containing 1 ng/mL EGF and 1 ng/mL bFGF. EdU was added 2 hr prior to fixation. Representative images of EdU (red) and DAPI (blue) staining in the culture treated with (A, left) vehicle (Ctrl) or (A, right) 30 μg/mL GLP are shown. The percentage of EdU + cells among total cells in the culture was determined (B). n = 3 independent experiments. (C–E) Adult hippocampal NPC were cultured in neurosphere-forming conditions in the presence of absence of GLP. Six days later, the number of neurospheres (C) and cells (D) were quantified for each condition. All neurospheres from each condition were collected, dissociated, and replated in the untreated culture medium. Six days later, the number of neurospheres was determined (E). n = 4 independent experiments. (F) Adult hippocampal NPC from APP/PS1 mice were cultured and treated the same as in (B), and the percentage of EdU + cells among total cells in the culture determined. n = 3 independent experiments. (G) Human iPSC-derived NPC cultures were treated with GLP of different concentration for 6 days in neurosphere-forming conditions. Neurospheres were quantified (n = 4 independent experiments). Quantifications are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 analyzed by one-way ANOVA followed by Fisher's protected least significant difference test. Scale bars, 100 μm. See also Figure S3 .

Article Snippet: Human iPSC-derived NPC were provided by IxCell Biotechnology and the monolayer cell culture was maintained in neural stem cell culturing medium (IxCell) in Matrigel (Corning)-coated dishes.

Techniques: In Vitro, Staining, Cell Culture, Derivative Assay, Concentration Assay

Journal: Cell Transplantation

Article Title: No Tumorigenicity of Allogeneic Induced Pluripotent Stem Cells in Major Histocompatibility Complex-matched Cynomolgus Macaques

doi: 10.1177/0963689721992066

Figure Lengend Snippet: Macaque iPSCs used in the present study show an immature phenotype with pluripotency. (A) Flow cytometric analysis of iPSCs. Lines indicate cells stained with antibodies against indicated molecules. Gray histograms indicate negative controls stained with isotype-matched control antibodies. *: Passage number. (B) HE-stained sections and immunohistochemistry of each teratoma derived from iPSCs in NOG mice after subcutaneous injection. The magnifications of the left panel and right panel of HE-stained sections are ×1 and ×40, respectively. The right panels are magnified images of the insets of the left panel. The magnifications of AFP (endoderm), αSMA (mesoderm), and β3-tubulin (ectoderm) are ×400. HE: hematoxylin and eosin; iPSC: induced pluripotent stem cell.

Article Snippet: The macaque iPSCs were passaged every 5–7 days by enzymatic digestion in CTK [0.6 mg/ml collagenase, 0.06% trypsin, 2.5 mM CaCl 2 , and 25% KnockoutTM Serum Replacement (KSR, Thermo Fisher Scientific, Waltham, MA, USA)] solution at 37°C for 5 min. Detached colonies containing MEFs were collected and cultured on the 0.1% gelatin-coated dish at 37°C for 15 to 30 min to remove MEFs again (attached–remove method).

Techniques: Staining, Control, Immunohistochemistry, Derivative Assay, Injection

Journal: Cell Transplantation

Article Title: No Tumorigenicity of Allogeneic Induced Pluripotent Stem Cells in Major Histocompatibility Complex-matched Cynomolgus Macaques

doi: 10.1177/0963689721992066

Figure Lengend Snippet: Macaque iPSCs form a teratoma in autologous transplantation but not in MHC-matched allogeneic transplantation. Monkeys #733 and #1497 were transplanted with CMF1/1 -1 cells in subcutaneous tissue of the shoulders and were observed for 12 weeks. (A) Schema of the transplantation. Autologous iPSCs, CMF1/1 -1 cells at passage 24, and iMSCs at passage 5 derived from CMF1/1 -1 cells were injected into subcutaneous tissue of the right shoulder (ˆ) and left shoulder (•) of macaque #733 with 50% Matrigel, respectively. (B) Schema of the transplantation. MHC-matched allogeneic iPSCs, CMF1/1 -1 cells at passage 41, and iMSCs at passage 4 derived from CMF1/1 -1 cells were respectively injected into subcutaneous tissue of the right shoulder (ˆ) and left shoulder (•) of macaque #1497 with 50% Matrigel. (C) The right shoulder of the iPSC transplantation site was swollen at 6 weeks after transplantation (circled), while the left shoulder of the iMSC transplantation site (arrows) was not swollen. (D) Ultrasonography revealed a heteroechoic mass (circled) in subcutaneous tissue on the scapula of the right shoulder but not that of the left shoulder 3 months after transplantation. (E) Neither shoulder of the cell transplantation site (arrows) was swollen 3 months after transplantation. (F) Ultrasonography revealed no mass in subcutaneous tissue in both shoulders 3 months after transplantation. (G) Gross appearance of the teratoma in the right shoulder of monkey #733. (H) HE-stained section of the teratoma in monkey #733. The magnification was ×40. (I) HE-stained section of biopsy specimens of the left shoulder of monkey #733 at 3 months after transplantation. The magnification was ×40. (J, K, and L) Immunohistochemistry of the teratoma. The magnifications of AFP (endoderm), αSMA (mesoderm), and β3-tubulin (ectoderm) were ×200, ×200 and ×400, respectively. HE: hematoxylin and eosin; iPSC: induced pluripotent stem cell; MHC: major histocompatibility complex.

Article Snippet: The macaque iPSCs were passaged every 5–7 days by enzymatic digestion in CTK [0.6 mg/ml collagenase, 0.06% trypsin, 2.5 mM CaCl 2 , and 25% KnockoutTM Serum Replacement (KSR, Thermo Fisher Scientific, Waltham, MA, USA)] solution at 37°C for 5 min. Detached colonies containing MEFs were collected and cultured on the 0.1% gelatin-coated dish at 37°C for 15 to 30 min to remove MEFs again (attached–remove method).

Techniques: Transplantation Assay, Derivative Assay, Injection, Staining, Immunohistochemistry, Immunopeptidomics