Transplantation Models to Characterize the Mechanisms of Stem Cell–Induced Islet Regeneration

Bell, Gillian I.; Seneviratne, Ayesh K.; Nasri, Grace N.; Hess, David A.

doi:10.1002/9780470151808.sc02b04s26

Cited by 3 publications

(3 citation statements)

References 44 publications

(50 reference statements)

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“…The effects of BM on human islet function observed in this study are consistent with most of reported results [ 20 , 33 ]. However, pure M and E cells improved islet β-cell function may be through mechanisms such as initiating angiogenesis [ 34 ] and preventing apoptosis [ 35 ] but less possibility of initiating β cells regeneration mechanisms supported by less significant β-cell regeneration relative transcription factor activations from human islet cocultured with M and E individually.…”

Section: Discussionsupporting

confidence: 93%

Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro

Luo

Xiong

Ravassard³

et al. 2015

BJMMR

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AimsAllogeneic bone marrow (BM) has been shown to support human islet survival and function in long-term culture by initiating human islet vascularization and β-cell regeneration. Various BM subpopulations may play different roles in human islet functions and survival. In this paper we investigated the effects of BM and its subpopulations, endothelial progenitor cells (E) and mesenchymal (M) cells on human islet's β-cell function and regeneration.Study DesignIsolation and identification of subpopulations from human bone marrow and culture with allogeneic human islet to investigate effects of different cell population on human islet function and regeneration.Place and Duration of StudyDepartment of Medicine, Center for Stem Cell & Diabetes Research, RWMC, Providence, RI, USA, between 2010 - 2014.MethodologyHuman islets were distributed from Integrated Islet Distribution Program (IIDP) and human bone marrow (BM) was harvested by Bone marrow transplantation center at Roger Williams Hospital. BM subpopulation was identified cell surface markers through Fluorescence-activated cell sorting, applied in flow cytometry (FACS), islet function was evaluated by human ELISA kit and β cell regeneration was evaluated by three methods of Cre-Loxp cell tracing, β cell sorting and RT-PCR for gene expression.ResultsFour different BM and seven different islet donates contributed human tissues. We observed islet β-cell having self regeneration capability in short term culture (3∼5 days) using a Cre-Loxp cell tracing. BM and its subtype E, M have similar benefits on β cell function during co-culture with human islet comparison to islet only. However, only whole BM enables to sustain the capability of islet β-cell self regeneration resulting in increasing β cell population while single E and M individual do not significantly affect on that. Mechanism approach to explore β-cell self regeneration by evaluating transcription factor expressions, we found that BM significantly increases the activations of β-cell regeneration relative transcription factors, the LIM homeodomain protein (Isl1), homologue to zebrafish somite MAF1 (MAFa), the NK-homeodomain factor 6.1 (NKX6.1), the paired box family factors 6 (PAX6), insulin promoter factor 1 (IPF1) and kinesin family member 4A (KIF4a).ConclusionThese results suggest that BM and its derived M and E cells enable to support human islet β-cell function. However, only BM can sustain the capability of β-cell self regeneration through initiating β-cell transcriptional factors but not individual E and M cells suggesting pure E and M cells less supportive for islet long-term survival in vitro.

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Section: Discussionsupporting

confidence: 93%

Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro

Luo

Xiong

Ravassard³

et al. 2015

BJMMR

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“…To induce hyperglycemia in non‐obese diabetic severe combined immune deficiency (NOD/SCID) mice, STZ (Sigma, germantown MD, USA) was administered intraperitoneally (35 mg/kg per day) daily for 5 days. For intrapancreatic (iPan) injections, mice were anesthetized; the pancreas and spleen exposed; a single dose of CM was microinjected (20 μl) into the splenic portion of the pancreas containing 4 μg or 8 μg total protein on day 10 (D10) as previously described . Control mice were injected with unconditioned media (basal media) and normoglycemic mice received citric acid buffer (CAB) vehicle instead of STZ.…”

Section: Methodsmentioning

confidence: 99%

Human Multipotent Stromal Cell Secreted Effectors Accelerate Islet Regeneration

et al. 2019

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Human multipotent stromal cells (hMSC) can induce islet regeneration after transplantation via the secretion of proteins that establish an islet regenerative niche. However, the identity of hMSCsecreted signals and the mechanisms by which pancreatic islet regeneration is induced remain unknown. Recently, mammalian pancreatic α-cells have been shown to possess considerable plasticity, and differentiate into β-like cells after near complete β-cell loss or overexpression of key transcriptional regulators. These studies have generated new excitement that islet regeneration during diabetes may be possible if we can identify clinically applicable stimuli to modulate these key regulatory pathways. Herein, we demonstrate that intrapancreatic-injection of concentrated hMSC-conditioned media (CM) stimulated islet regeneration without requiring cell transfer. hMSC CM-injection significantly reduced hyperglycemia, increased circulating serum insulin concentration, and improved glucose tolerance in streptozotocin-treated mice. The rate and extent of endogenous β-cell mass recovery was dependent on total protein dose administered and was further augmented by the activation of Wnt-signaling using GSK3-inhibition during CM generation. Intrapancreatic hMSC CM-injection immediately set in motion a cascade of regenerative events that included the emergence of proliferating insulin + clusters adjacent to ducts, NKX6.1 expression in glucagon + cells at days 1-4 suggesting the acquisition of β-cell phenotype by α-cells, and accelerated β-cell maturation with increased MAFA-expression for >1 month postinjection. Discovery and validation of islet regenerative hMSC-secreted protein may lead to the development of cell-free regenerative therapies able to tip the balance in favor of β-cell regeneration versus destruction during diabetes. STEM CELLS 2019;37:516-528 SIGNIFICANCE STATEMENTCell-based therapies using human multipotent stromal cells (hMSC) to dampen autoimmunity while inducing islet regeneration represents a promising approach to treat diabetes. However, poor survival and retention of transplanted hMSC in the damaged pancreas complicates clinical translation. Here, it is demonstrated that hMSC secrete an array of islet-regenerative proteins and intrapancreatic delivery of hMSC-conditioned media (CM) could stimulate islet regeneration without the need for cell transfer. Wnt-pathway stimulated hMSC CM-injection set in motion a cascade of events consistent with the emergence of functional islets. Discovery of hMSC-secreted, islet-regenerative effectors may lead to the development of cell-free therapies to combat diabetes.

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“…Immunodeficient NOD/SCID mice (Jackson Labs) were intraperitoneally‐injected (i.p.) with 35 mg/kg streptozotocin (STZ) (Sigma‐Aldrich) for 5 consecutive days, as previously described . Mice reaching hyperglycemia on day 10 (15–25 mmol/L) were sublethally irradiated (300 cGy) and transplanted by tail vein injection with PBS or MSCs (5 × 10 5 ).…”

Section: Methodsmentioning

confidence: 99%

Brief Report: Elastin Microfibril Interface 1 and Integrin-Linked Protein Kinase Are Novel Markers of Islet Regenerative Function in Human Multipotent Mesenchymal Stromal Cells

et al. 2016

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Multipotent mesenchymal stromal cell (MSC) transplantation is proposed as a novel therapy for treating diabetes by promoting the regeneration of damaged islets. The clinical promise of such treatments may be hampered by a high degree of donor-related variability in MSC function and a lack of standards for comparing potency. Here, we set out to identify markers of cultured human MSCs directly associated with islet regenerative function. Stromal cultures from nine separate bone marrow donors were demonstrated to have differing capacities to reduce hyperglycemia in the NOD/SCID streptozotocin-induced diabetic model. Regenerative (R) and nonregenerative (NR) MSC cultures were directly compared using isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics. A total of 1,410 proteins were quantified resulting in the identification of 612 upregulated proteins and 275 downregulated proteins by 6 1.2-fold in R-MSC cultures. Elastin microfibril interface 1 (EMILIN-1), integrinlinked protein kinase (ILK), and hepatoma-derived growth factor (HDGF) were differentially expressed in R-MSCs, and Ingenuity Pathway Analyses revealed each candidate as known regulators of integrin signaling. Western blot validation of EMILIN-1, ILK, and HDGF not only showed significantly higher abundance levels in R-MSCs, as compared with NR-MSCs, but also correlated with passage-induced loss of islet-regenerative potential. Generalized estimating equation modeling was applied to examine the association between each marker and blood glucose reduction. Both EMILIN-1 and ILK were significantly associated with blood glucose lowering function in vivo. Our study is the first to identify EMILIN-1 and ILK as prospective markers of islet regenerative function in human MSCs. STEM CELLS 2016;34:2249-2255 SIGNIFICANCE STATEMENTThe vast majority of current stem cell based clinical trials are testing the therapeutic capacity of multipotent mesenchymal stem cells for treating a wide range of diseases. One of the critical issues plaguing the progress of these trials is a lack of suitable and reliable methods for measuring the potency of these stem cells. This study uncovers two new potential proteins whose abundance in human bone marrow mesenchymal stem cell cultures provides an accurate indication of their potency for repairing damaged pancreatic islets, as would be found patients suffering from Type I diabetes. We predict these proteins to impact the development of novel stem cell products for treating pancreatic diseases.

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Transplantation Models to Characterize the Mechanisms of Stem Cell–Induced Islet Regeneration

Cited by 3 publications

References 44 publications

Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro

Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro

Human Multipotent Stromal Cell Secreted Effectors Accelerate Islet Regeneration

Brief Report: Elastin Microfibril Interface 1 and Integrin-Linked Protein Kinase Are Novel Markers of Islet Regenerative Function in Human Multipotent Mesenchymal Stromal Cells

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