Journal: JCI Insight

Article Title: Modeling immune responses to autologous and allogeneic human stem cell–derived islet grafts in vivo

doi: 10.1172/jci.insight.200738

Figure Lengend Snippet: ( A and B ) Human immune system reconstitution in cohort 1 ( A ) and 2 ( B ); human chimerism is measured as the percentage of human CD45 + cells among total CD45 + cells. Each symbol represents an individual animal. In A , a hollow triangle represents an allogeneic mouse not injected with polyclonal T cells. ( C ) CD34 – nonadherent fetal liver cells reprogramming into iPSCs using Klf4, Oct3/4, Sox2, and c-Myc transcription factors delivered via the nonintegrating Sendai virus, with examples of iPSC colonies. Scale bar: 200 µm. ( D ) iPSC-to-β cell differentiations performed in the study. ( E ) Circulating C-peptide levels in autologous (cohort 1 and 2) and allogeneic grafts. In autologous cohort 1 and 2, n = 2/3 and n = 3/12 mice were injected with IAR-TCR + T cells that did not infiltrate the grafts, respectively, and n = 5/6 allogeneic mice were injected with additional in vitro activated polyclonal T cells from a mouse with the same HIS. In E , the asterisk indicates the level of significance (* P < 0.05) when comparing the percentage of circulating C-peptide in allogeneic and autologous cohort 2 at weeks 12 and 15 after β cell transplantation. ( F ) Correlation between percentage of C-peptide + cells in islet before transplant and circulating human C-peptide levels 12 and 15 weeks later. In A , B , and E , data show the mean ± SEM. In F , different groups are represented with the same symbols as E, and each symbol represents the mean of the group. Statistical analysis was performed using a mixed-effects model with Geisser-Greenhouse correction and Tukey’s post hoc test for E and a simple linear regression for F . C-1, cohort 1; C-2, cohort 2.

Article Snippet: Circulating human C-peptide was measured on 20 μL of plasma, isolated from whole blood, using the STELLUX Chemi Human C-peptide ELISA kit (Alpco).

Techniques: Injection, Virus, In Vitro, Transplantation Assay

Journal: JCI Insight

Article Title: Modeling immune responses to autologous and allogeneic human stem cell–derived islet grafts in vivo

doi: 10.1172/jci.insight.200738

Figure Lengend Snippet: ( A ) Endocrine cell populations found in the grafts, represented as percentage of total endocrine cells. ( B and C ) Comparison of C-peptide + ( B ) and C-peptide + NKX6.1 + cells ( C ) among cells found in the islet region between autologous (cohort 1, cohort 2) and allogeneic grafts. ( D ) Representative images of endocrine cell subpopulations in autologous grafts from cohort 1. Scale bar: 100 µm. ( E ) Representative images of endocrine cell subpopulations and CD57 expression in the grafts of autologous cohort 2. Scale bar: 100 µm. ( F ) Quantification of CD57 + cells in the grafts. ( G ) Quantification of CD57 expression among SC-islet cell subtypes. For D and E , markers used for analysis are indicated in the figure. In autologous cohorts 1 and 2, n = 2 and n = 3 mice were injected with IAR-TCR + T cells, respectively; n = 5 allogeneic mice were injected with additional in vitro activated polyclonal T cells (a hollow triangle represents the allogeneic mouse not injected with additional polyclonal T cells, and autologous mice injected with IAR-TCR + T cells are represented as solid circles). Mean ± SEM; 1-way ANOVA/Tukey’s post hoc test was used for B , C , and F . * P < 0.05, ** P < 0.01. C-1, cohort 1; C-2, cohort 2; SST, somatostatin.

Article Snippet: Circulating human C-peptide was measured on 20 μL of plasma, isolated from whole blood, using the STELLUX Chemi Human C-peptide ELISA kit (Alpco).

Techniques: Comparison, Expressing, Injection, In Vitro

Journal: JCI Insight

Article Title: Modeling immune responses to autologous and allogeneic human stem cell–derived islet grafts in vivo

doi: 10.1172/jci.insight.200738

Figure Lengend Snippet: ( A ) Quantification of CD45 + cell proliferation in autologous and allogeneic grafts. ( B ) Example of CD45 + cell proliferation in allogeneic grafts. Scale bar: 100 µm. ( C ) Quantification of CD3 + cell proliferation in autologous and allogeneic grafts. ( D and E ) Quantification of glucagon + α ( D ) and C-peptide + β ( E ) cell proliferation in autologous and allogeneic grafts. ( F ) Example of α and β cell proliferation in allogeneic grafts. Scale bar: 100 µm. ( G ) Example of CD45 + α and β cell proliferation in autologous grafts. n = 5 allogeneic mice were injected with additional in vitro activated polyclonal T cells (a hollow triangle represents the allogeneic mouse not injected with polyclonal T cells). Scale bar: 100 µm. Panel in F represents a lower-magnification view of the area shown in , visualized with different markers. Data shown as mean ± SEM; statistical analysis was performed with an unpaired 2-tailed t test for A and C – E . * P < 0.05. C-1, cohort 1; C-2, cohort 2.

Article Snippet: Circulating human C-peptide was measured on 20 μL of plasma, isolated from whole blood, using the STELLUX Chemi Human C-peptide ELISA kit (Alpco).

Techniques: Injection, In Vitro