TITLE:
Bone marrow increases human islets insulin positive cells in co-culture: quantification with flow cytometry
AUTHORS:
Zhengke Wang, Fang Xiong, Mary Hassani, John Z. Q. Luo, LuGuang Luo
KEYWORDS:
Bone marrow; Human Islets; Flow Cytometry
JOURNAL NAME:
Journal of Diabetes Mellitus,
Vol.1 No.4,
November
8,
2011
ABSTRACT: 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.