Abstract:In this study, we have established a new strategy increasing human islet longevity utilizing allogeneic whole bone marrow (BM) co-cultured with human islets. The cultured islets' function and survival have been evaluated by analysis of insulin secretion in response to high-glucose-challenge, morphological evaluation of cell growth. Human islet only culture failed to reveal evidence of long term survival, growth or function in terms of insulin release or insulin response to glucose challenge. These results indi… Show more
“…Human islet tissue from normal donors was obtained through the Islet Resource Centers (ICRs) Basic Science Islet Distribution Program from the Human Islet Laboratories at the University of Pennsylvania (Philadelphia, PA, USA), Massachusetts General Hospital (Boston, MA, USA), City of Hope National Medical Center (Duarte, CA, USA), University of Miami, School of Medicine, Diabetes Research Institute (Miami, FL, USA). The use of these cells is approved by the Institutional Review Board (IRB) at Roger Williams Hospital and the ICR Committee [9].…”
Section: Tissue and Cell Culturementioning
confidence: 99%
“…Trypan blue (Gibco, 15250-061) staining was used to assess cell viability. The use of these cells is approved by the Institutional Review Board (IRB) at Roger Williams Hospital and the ICR Committee [9].…”
Section: Tissue and Cell Culturementioning
confidence: 99%
“…Our previous studies have indicated that bone marrow can maintain β cell function by evaluating insulin secretion even after 200 days of in vitro culture [9]. Exploration of mechanisms how BM support human islets in long term suggests that BM cultured human islet may increase β-cell population in islet during culture.…”
We have previously demonstrated that allogeneic human bone marrow (BM) supports human islet function and longevity in vitro. We hypothesize that BM supporting human islets may include to increase β-cell in cultured islets. In this study, we developed a method to quantify insulin-producing β cells from cultured islets by using immunofluorescent staining and flow cytometry analysis to explore this possibility. The results show that human islets cocultured with BM for 39 days contained a significantly higher number of insulin-positive β cells (42.3% ± 4.5%) compared to the islet-only cultures (1.15% ± 0.78%), and increased insulin release levels evaluated by ELISA is consistent with increased β cells in same culture condition. Human islet culture with BM significantly increase β-cells while islet only culture lost β-cells in same culture period supports the possibility of BM increasing β-cells in cultured islets.
“…Human islet tissue from normal donors was obtained through the Islet Resource Centers (ICRs) Basic Science Islet Distribution Program from the Human Islet Laboratories at the University of Pennsylvania (Philadelphia, PA, USA), Massachusetts General Hospital (Boston, MA, USA), City of Hope National Medical Center (Duarte, CA, USA), University of Miami, School of Medicine, Diabetes Research Institute (Miami, FL, USA). The use of these cells is approved by the Institutional Review Board (IRB) at Roger Williams Hospital and the ICR Committee [9].…”
Section: Tissue and Cell Culturementioning
confidence: 99%
“…Trypan blue (Gibco, 15250-061) staining was used to assess cell viability. The use of these cells is approved by the Institutional Review Board (IRB) at Roger Williams Hospital and the ICR Committee [9].…”
Section: Tissue and Cell Culturementioning
confidence: 99%
“…Our previous studies have indicated that bone marrow can maintain β cell function by evaluating insulin secretion even after 200 days of in vitro culture [9]. Exploration of mechanisms how BM support human islets in long term suggests that BM cultured human islet may increase β-cell population in islet during culture.…”
We have previously demonstrated that allogeneic human bone marrow (BM) supports human islet function and longevity in vitro. We hypothesize that BM supporting human islets may include to increase β-cell in cultured islets. In this study, we developed a method to quantify insulin-producing β cells from cultured islets by using immunofluorescent staining and flow cytometry analysis to explore this possibility. The results show that human islets cocultured with BM for 39 days contained a significantly higher number of insulin-positive β cells (42.3% ± 4.5%) compared to the islet-only cultures (1.15% ± 0.78%), and increased insulin release levels evaluated by ELISA is consistent with increased β cells in same culture condition. Human islet culture with BM significantly increase β-cells while islet only culture lost β-cells in same culture period supports the possibility of BM increasing β-cells in cultured islets.
“…Concomitant transplantation of islets with BM stem cells, vascular endothelial cells, endothelial progenitor cells (EPCs) and MSCs creates a suitable niche for promotion of revascularization and enhancement of grafted islet survival and function (123,124,125,126).…”
Section: Enhancement Of Islet Vasculaturementioning
Over the past decades, tremendous efforts have been made to establish pancreatic islet transplantation as a standard therapy for type 1 diabetes. Recent advances in islet transplantation have resulted in steady improvements in the 5-year insulin independence rates for diabetic patients. Here we review the key challenges encountered in the islet transplantation field which include islet source limitation, sub-optimal engraftment of islets, lack of oxygen and blood supply for transplanted islets, and immune rejection of islets. Additionally, we discuss possible solutions for these challenges.
“…On the other hand, most studies have shown beneficial effects of cellular and cytokine interventions on islet remodeling and recovery from injury without significant conversion to produce insulin (8,12,19,33,39,40,42,45,66,68).…”
We have recently reported that small-sized bone marrow cells (BMCs) isolated by counterflow centrifugal elutriation and depleted of lineage markers (Fr25lin − ) have the capacity to differentiate and contribute to regeneration of injured islets. In this study, we assess some of the characteristics of these cells compared to elutriated hematopoietic progenitors (R/O) and whole BMCs in a murine model of streptozotocin-induced chemical diabetes. The GFP bright CD45+ progeny of whole BMCs and R/O progenitors progressively infiltrate the pancreas with evolution of donor chimerism; are found at islet perimeter, vascular, and ductal walls; and have a modest impact on islet recovery from injury. In contrast, Fr25lin− cells incorporate in the islets, convert to GFP dim CD45 − PDX-1 + phenotypes, produce proinsulin, and secrete insulin with significant contribution to stabilization of glucose homeostasis. The elutriated Fr25lin − cells express low levels of CD45 and are negative for SCA-1 and c-kit, as removal of cells expressing these markers did not impair conversion to produce insulin. BMCs mediate two synergistic mechanisms that contribute to islet recovery from injury: support of islet remodeling by hematopoietic cells and neogenesis of insulin-producing cells from stem cells.
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