Islet Cell Purification and Characterization

Islet cell preparations were characterized by their number of beta cells after purification and after a 1- to 5-day culture period [determined after a minimum of two washing cycles to eliminate damaged cells and cell debris, by combining cell number (nuclear count assay in duplicate samples; NucleoCounter YC-100; ChemoMetec, Allerød, Denmark) and percentage of insulin positive cells (immunocytochemistry; >10³ cells counted)] [7 (link), 22 (link)]. Yield immediately post purification was also expressed as islet equivalent (IEQ), calculated using a volume based method after dithizone (DTZ) staining [23 (link), 24 ]. Only preparations with these data available after purification were included for analysis. Cellular insulin content was determined and expressed as μg insulin / 10⁶ beta cells [22 (link)].

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De Paep D.L., Van Hulle F., Ling Z., Vanhoeij M., Pirenne J., Keymeulen B., Pipeleers D, & Jacobs-Tulleneers-Thevissen D. (2021). Lower beta cell yield from donor pancreases after controlled circulatory death prevented by shortening acirculatory warm ischemia time and by using IGL-1 cold preservation solution. PLoS ONE, 16(5), e0251055.

Publication 2021

NucleoCounter YC-100

Assay Cellular Dithizone Immunocytochemistry Insulin Insulin cells Islet cell Staining method

Corresponding Organization : KU Leuven

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Variable analysis

independent variables

Culture period duration (1- to 5-day)

dependent variables

Number of beta cells after purification
Number of beta cells after culture period
Percentage of insulin positive cells
Islet equivalent (IEQ) yield immediately post purification
Cellular insulin content (μg insulin / 10^6 beta cells)

control variables

Minimum of two washing cycles to eliminate damaged cells and cell debris

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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