GM-CSF treated F4/80+ BMCs improve murine hind limb ischemia similar to M-CSF differentiated macrophages.

Novel cell therapy is required to treat critical limb ischemia (CLI) as many current approaches require repeated aspiration of bone marrow cells (BMCs). The use of cultured BMCs can reduce the total number of injections required and were shown to induce therapeutic angiogenesis in a murine model of hind limb ischemia. Blood flow recovery was significantly improved in mice treated with granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent BMCs that secreted inflammatory cytokines. Angiogenesis, lymphangiogenesis, and blood flow recovery ratio were significantly higher in the GM-CSF-cultured F4/80+ macrophage (GM-Mø)-treated group compared with controls. Furthermore, Foxp3+ cell numbers and tissue IL-10 concentrations were significantly increased compared with controls. There was no significant difference in blood flow recovery between GM-Mø and M-CSF-cultured F4/80+ macrophages (M-Mø). Thus, GM-Mø were associated with improved blood flow in hind limb ischemia similar to M-Mø. The selective methods of culturing and treating GM-Mø cells similar to M-Mø cells could be used clinically to help resolve the large number of cells required for BMC treatment of CLI. This study demonstrates a novel cell therapy for CLI that can be used in conjunction with conventional therapy including percutaneous intervention and surgical bypass

Expansion of transplanted islets in mice by co-transplantation with adipose tissue-derived mesenchymal stem cells

The shortage of donor islets is a significant obstacle for widespread clinical application of pancreatic islet transplantation. To investigate whether adipose tissue-derived mesenchymal stem cells (ADSCs) induce expansion of transplanted islets, we performed co-transplantation experiments in a mouse model. Streptozotosin (STZ)-induced diabetic mice transplanted with 50 syngeneic islets remained hyperglycemic. However, hyperglycemia was ameliorated gradually when 50 islets were co-transplanted with ADSCs but not separately grafted into the contralateral kidney. Insulin and proinsulin contents of 120-day grafts containing 50 islets co-transplanted with ADSCs were significantly increased compared with those of 50 isolated islets. The Ki67-positive ratios in islets of the naïve pancreas, at 30 and 120 days grafts were 0.23%, 2.12%, and 1.52%, respectively. Ki67-positive cells were predominantly Pdx1+ and insulin+ cells. These results demonstrate that co-transplantation with ADSCs induces proliferation of transplanted islets in mice, suggesting a potential solution for the low efficiency of islet transplantation

vWF and LYVE-1 staining in M-Mø and GM-Mø treated ischemic hind limbs.

<p>Representative photomicrographs (original magnification ×400). vWF positive and LYVE-1 positive cells are stained green and red, respectively (***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05).</p

Difference in Neutropenia due to Administration Schedule of TAS-102

TAS-102 significantly improves overall survival in patients with metastatic colorectal cancer. The most common adverse event of TAS-102 is bone marrow suppression, which leads to neutropenia. The incidence of neutropenia is high, and there is no known effective prevention method. Furthermore, the administration method of TAS-102 is complicated. We reported that neutropenia could be avoided by changing to a simple administration method of TAS-102

Long-Term Tracking of the Effects of Colostrum-Derived <i>Lacticaseibacillus rhamnosus</i> Probio-M9 on Gut Microbiota in Mice with Colitis-Associated Tumorigenesis

Intestinal bacteria play important roles in the progression of colitis-associated carcinogenesis. Colostrum-derived Lacticaseibacillus rhamnosus Probio-M9 (Probio-M9) has shown a protective effect in a colitis-associated cancer (CAC) model, but detailed metagenomic analysis had not been performed. Here, we investigated the preventive effects of the probiotic Probio-M9 on CAC-model mice, tracking the microbiota. Feces were obtained at four time points for evaluation of gut microbiota. The effect of Probio-M9 on tight junction protein expression was evaluated in co-cultured Caco-2 cells. Probio-M9 treatment decreased the number of tumors as well as stool consistency score, spleen weight, inflammatory score, and macrophage expression in the CAC model. Probio-M9 accelerated the recovery of the structure, composition, and function of the intestinal microbiota destroyed by azoxymethane (AOM)/dextran sulfate sodium (DSS) by regulating key bacteria (including Lactobacillus murinus, Muribaculaceae bacterium DSM 103720, Muribaculum intestinale, and Lachnospiraceae bacterium A4) and pathways from immediately after administration until the end of the experiment. Probio-M9 co-culture protected against lipopolysaccharide-induced impairment of tight junctions in Caco-2 cells. This study provides valuable insight into the role of Probio-M9 in correcting gut microbiota defects associated with inflammatory bowel disease carcinogenesis and may have clinical application in the treatment of inflammatory carcinogenesis

Flow cytometric analysis of F4/80⁺ macrophages from M-CSF- or GM-CSF- cultured BMCs.

<p>Data is representative of three independent experiments.</p

Expression of cytokine mRNA, and IL-10 protein levels in M-CSF- or GM-CSF-cultured BMCs.

<p>(A) The expression of proinflammatory cytokines (TNF-alpha, IL-6, IL-1beta, IL-12a, and IL12b) and anti-inflammatory cytokine (IL-10) mRNA in M-CSF- or GM-CSF-cultured BMCs. (B) IL-10 concentration in the supernatant of M-CSF- or GM-CSF-cultured BMCs. Data are expressed as mean ± SEM, ***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05.</p

Recovery of blood flow in ischemic limbs treated with M-CSF-cultured BMCs and GM-CSF-cultured BMCs.

<p>Representative laser Doppler perfusion images taken at indicated intervals for the PBS control group, M-CSF-cultured BMCs group and GM-CSF-cultured BMC group. Data are expressed as mean ± SEM, n = 5–7/group, **<i>P</i><0.01, *<i>P</i><0.05.</p

Foxp3 expression in M-Mø or GM-Mø treated ischemic hind limbs and the recovery of blood flow in ischemic limbs treated with IL-10.

<p>(A) Photomicrographs show representative DAB staining for Foxp3 (scale bar = 50 µm). The number of Foxp3 positive cells was counted in five random images for each mouse. The thigh muscle of each group was isolated 7 days after the induction of ischemia. (B) IL-10 levels in thigh muscle after limb ischemia at 7 days after intramuscular injection of indicated cells (data are expressed as mean ± SEM, n = 3 group, ***<i>P</i><0.001, **<i>P</i><0.01, *<i>P</i><0.05). (C) Recovery of blood flow in ischemic limbs treated IL-10. Representative laser doppler perfusion images were taken at indicated intervals for the PBS control groups and IL-10 treated groups. Data are expressed as mean ± SEM, n = 5–7/group.</p

Recovery of blood flow in ischemic limbs treated with M-Mø and GM-Mø.

<p>Representative laser Doppler perfusion images were taken at indicated intervals for the PBS control group, M-Mø group, and GM-Mø group. Data are expressed as mean ± SEM, n = 5 group, **<i>P</i><0.01, *<i>P</i><0.05.</p