Bone marrow increases human islets insulin positive cells in co-culture quantification with flow cytometry英文文献资料.docVIP

Vol.1, No.4, 109-117 (2011) Journal of Diabetes Mellitus doi:10.4236/jdm.2011.14015 Bone marrow increases human islets insulin positive cells in co-culture: quantification with flow cytometry Zhengke Wang , Fang Xiong , Mary Hassani , John Z. Q. Luo , LuGuang Luo 1 1 1 1,2 1* 1 2 Center for Stem Cell Biology, Roger Williams Hospital, Providence, USA; Corresponding Author: Lluo@ * Alpert Medical School, Brown University, Providence, USA. Received 30 August 2011; revised 4 October 2011; accepted 15 October 2011. ABSTRACT results in a significant loss of structural integrity within the islets and a decline in glucose-stimulated insulin re- lease [6-8]. We have previously demonstrated that allogene- ic human bone marrow (BM) supports human islet function and longevity in vitro. We hypoth- esize that BM supporting human islets may incl- ude to increase β-cell in cultured islets. In this study, we developed a method to quantify insu- lin-producing β cells from cultured islets by us- ing immunofluorescent staining and flow cyto- metry 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 eva- luated by ELISA is consistent with increased β cells in same culture condition. Human islet cul- ture with BM significantly increase β-cells while islet only culture lost β-cells in same culture pe- riod supports the possibility of BM increasing β-cells in cultured islets. Our previous studies have indicated that bone marrow can maintain β cell function by evaluating insulin secre- tion even after 200 days of in vitro culture [9]. Explora- tion of mechanisms