Journal: Science Advances
Article Title: 3D bioprinting of collagen-based high-resolution internally perfusable scaffolds for engineering fully biologic tissue systems
doi: 10.1126/sciadv.adu5905
Figure Lengend Snippet: ( A ) Schematic design and 3D printer machine pathing G-code of a dual parallel channel multi-material CHIPS with pancreatic vascular cell bioink (MIN6, HUVEC, and MSC) regions surrounding both sides of the channels. ( B ) Pancreatic CHIPS FRESH printed and visualized via bright-field stereomicroscope (inset) and 3D confocal fluorescence imaging of the optically cleared pancreatic scaffold after 12 days of static culture. ( C ) XY midplane view from confocal fluorescence image of the 12-day statically cultured pancreatic CHIPS following 3D vascular network segmentation for quantification of network diameter and density within the migratory zones. ( D ) Example confocal fluorescence images revealing additional cell migration into the acellular regions of the CHIPS beneath the cellular regions guided by the printed collagen filaments. ( E ) XY midplane projection view from 3D confocal fluorescence imaging of the optically cleared 12-day VAPOR perfused pancreatic CHIPS. ( F ) Graphic illustration and example ROIs depicting evidence of early MIN6 pancreatic bud and microlumen formation with actin (green) and insulin (magenta) fluorescence images from ROIs 2 and 3 in (E). ( G ) Graphic illustration and quantification for insulin secretion ELISA assay from 1.5-hour glucose-stimulated [ratio of high glucose (HG) to low glucose (LG)] insulin secretion experiment between 12-day static and perfusion cultured pancreatic CHIPS (means ± SD; ** P < 0.01 for N = 3 static tissues; N = 2 perfused tissues, unpaired t test).
Article Snippet: MIN6 (mouse insulinoma cell line) (Addexbio, C0018008) were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM, Gibco, 11-965-092) with 15% fetal bovine serum (v/v), 2 mM sodium pyruvate, 20 mM Hepes, and 0.05 mM β-mercaptoethanol, using passages 9 to 15.
Techniques: Fluorescence, Imaging, Cell Culture, Migration, Enzyme-linked Immunosorbent Assay
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![( A ) Schematic design and 3D printer machine pathing G-code of a dual parallel channel multi-material CHIPS with pancreatic vascular cell bioink <t>(MIN6,</t> HUVEC, and MSC) regions surrounding both sides of the channels. ( B ) Pancreatic CHIPS FRESH printed and visualized via bright-field stereomicroscope (inset) and 3D confocal fluorescence imaging of the optically cleared pancreatic scaffold after 12 days of static culture. ( C ) XY midplane view from confocal fluorescence image of the 12-day statically cultured pancreatic CHIPS following 3D vascular network segmentation for quantification of network diameter and density within the migratory zones. ( D ) Example confocal fluorescence images revealing additional cell migration into the acellular regions of the CHIPS beneath the cellular regions guided by the printed collagen filaments. ( E ) XY midplane projection view from 3D confocal fluorescence imaging of the optically cleared 12-day VAPOR perfused pancreatic CHIPS. ( F ) Graphic illustration and example ROIs depicting evidence of early <t>MIN6</t> pancreatic bud and microlumen formation with actin (green) and insulin (magenta) fluorescence images from ROIs 2 and 3 in (E). ( G ) Graphic illustration and quantification for insulin secretion ELISA assay from 1.5-hour glucose-stimulated [ratio of high glucose (HG) to low glucose (LG)] insulin secretion experiment between 12-day static and perfusion cultured pancreatic CHIPS (means ± SD; ** P < 0.01 for N = 3 static tissues; N = 2 perfused tissues, unpaired t test).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7336/pmc12017336/pmc12017336__sciadv.adu5905-f7.jpg)