Luo Lu Guang, Xiong Fang, Ravassard Philippe, Luo John Zq
Department of Medicine/Research, Roger Williams Medical Center, Boston University, USA.
Department of Molecular Biology, ICM, Biotechnology & Biotherapy Group 47 Bd de Hospital, Paris France.
Br J Med Med Res. 2015;8(7):576-587. doi: 10.9734/BJMMR/2015/17536. Epub 2015 May 18.
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.
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.
Department of Medicine, Center for Stem Cell & Diabetes Research, RWMC, Providence, RI, USA, between 2010 - 2014.
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 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.
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 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).
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 .
异体骨髓已被证明可通过启动人胰岛血管生成和β细胞再生，在长期培养中支持人胰岛的存活和功能。各种骨髓亚群可能在人胰岛功能和存活中发挥不同作用。在本文中，我们研究了骨髓及其亚群、内皮祖细胞（E）和间充质（M）细胞对人胰岛β细胞功能和再生的影响。
从人骨髓中分离和鉴定亚群，并与异体人胰岛共培养，以研究不同细胞群体对人胰岛功能和再生的影响。
美国罗德岛州普罗维登斯市RWMC干细胞与糖尿病研究中心医学部，2010年至2014年。
人胰岛来自综合胰岛分配计划（IIDP），人骨髓由罗杰·威廉姆斯医院骨髓移植中心采集。通过荧光激活细胞分选，利用细胞表面标志物鉴定骨髓亚群，应用于流式细胞术（FACS），用人ELISA试剂盒评估胰岛功能，通过三种Cre-Loxp细胞追踪、β细胞分选和基因表达RT-PCR方法评估β细胞再生。
四个不同的骨髓和七个不同的胰岛供体提供了人体组织。我们使用Cre-Loxp细胞追踪观察到胰岛β细胞在短期培养（3至5天）中有自我再生能力。与仅培养胰岛相比，骨髓及其亚型E、M在与人胰岛共培养期间对β细胞功能有类似益处。然而，只有全骨髓能够维持胰岛β细胞自我再生的能力，从而增加β细胞数量，而单个E和M个体对此没有显著影响。通过评估转录因子表达来探索β细胞自我再生的机制方法，我们发现骨髓显著增加了β细胞再生相关转录因子的激活，即LIM同源域蛋白（Isl1）、斑马鱼体节MAF1（MAFa）的同源物、NK同源域因子6.1（NKX6.1）、配对盒家族因子6（PAX6）、胰岛素启动子因子1（IPF1）和驱动蛋白家族成员4A（KIF4a）。
这些结果表明，骨髓及其衍生的M和E细胞能够支持人胰岛β细胞功能。然而，只有骨髓能够通过启动β细胞转录因子来维持β细胞自我再生的能力，而单个E和M细胞则不能，这表明纯E和M细胞对胰岛长期存活的支持作用较小。
