FIG. 8.

ADMSC therapy promotes long-term reversion of hyperglycemia in diabetic NOD mice. ADMSCs (1 × 10⁶) were administrated by intraperitoneal injection in female NOD diabetic mice (blood glucose >240 mg/dL by two consecutive measurements) on days 0, 7, and 14. A: Blood glucose (mg/dL) was measured once a week after treatment under nonfasting conditions for 12 weeks. The gray region denotes blood glucose values <240 mg/dL. Average ± SEM of blood glucose levels in the initiation of treatment is shown and did not differ between groups (P = NS). The reversion of hyperglycemia could be maintained for 8 weeks in seven of nine of the ADMSC-treated mice. B: The weekly mean blood glucose levels of responder mice were significantly reduced compared with the untreated group from 2 to 12 weeks. Normoglycemic levels were detected in 57% (four of seven) of responder mice with no insulin administration. C: Histological analysis of the pancreas by H-E and immunohistochemistry for insulin expression, respectively. Original magnification ×200. D: The quantification of mononuclear cellular infiltrate (or insulitis score) in the pancreatic islets was determined by counting at least 80 islets/experimental group from at least five mice/group using the following classification: 0 (no cellular infiltration), 0–25, 25–50, >50, and 100% of cell infiltration in each islet. The frequency of Tregs was analyzed by flow cytometry in cells obtained from the PLNs of responder mice. The cells were stained for surface markers CD4 and CD25 and afterward for transcription factor Foxp3. E: Dot plots represent the frequency of CD4⁺Foxp3⁺ cells at the CD4⁺ cell gate. F and G: Frequency of CD4⁺Foxp3⁺ and CD4⁺CD25⁺Foxp3⁺ T cells in the CD4⁺ cell gate, respectively, for each group. H: Dot plots represent the frequency of CD4⁺Foxp3⁺Helios⁺ cells at the CD4⁺ cell gate. I and J: Frequency of CD4⁺Foxp3⁺Helios⁻ and CD4⁺Foxp3⁺Helios⁺ in the CD4⁺ cell gate, respectively. Data are average ± SEM. *P < 0.05; **P < 0.01. (A high-quality color representation of this figure is available in the online issue.)