Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro.
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Date
2015
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Abstract
Aims: Allogeneic 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 Design: Isolation 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 Study: Department of Medicine, Center for Stem Cell & Diabetes
Research, RWMC, Providence, RI, USA, between 2010 - 2014.
Methodology: Human 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 Fluorescenceactivated
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.
Results: Four 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 coculture
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).
Conclusion: These 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.
Description
Keywords
Allogeneic bone marrow, mesenchymal cells, endothelia cells, human Islet, β-cell regeneration
Citation
Luo Lu Guang, Xiong Fang, Ravassard Philippe, Luo John ZQ. Human Bone Marrow Subpopulations Sustain Human Islet Function and Viability In vitro. British Journal of Medicine and Medical Research. 2015; 8(7): 576-587.