H9 hEBs (formed under -ROCK/-spin condition) differentiated into pancreatic lineage at 18 days when treated with the pancreatic differentiation protocol.

<p>(<b>a</b>) RT-PCR analysis demonstrated expression of key pancreatic lineage-specific genes. (<b>b</b>) Over 90% of the cells were insulin-positive, as evidenced by positive staining for dithizone (DTZ) in dark red. (<b>c</b>) Immunostaining of the cells revealed co-localization (in yellow) of insulin and C-peptide. Cell nuclei were stained in blue with DAPI.</p

H9 hESCs formed hEBs in the microwells at a cell seeding density of 25,000 hESCs per microwell.

<p>(<b>a</b>) Freshly extracted hEBs formed under four different conditions in microwells after 24 hrs of incubation. (<b>b</b>) A closer look at hEBs formed under -ROCK/-spin condition after 24 hrs of incubation before extraction indicated the presences of a compact core (arrows) and a loosely-aggregated corona. (<b>c</b>) Live-dead staining of the formed hEBs under -ROCK/-spin condition after 24 hrs of incubation before transfer to suspension culture indicated high viability (>90%) of hESCs both at the core and the corona (SYTO 10 green-fluorescent nucleic acid stain for all cells; DEAD Red (ethidium homodimer-2) nucleic acid stain for dead cells). (<b>d</b>) Confocal microscopic images of live/dead staining of freshly extracted hEBs (formed under -ROCK/-spin condition) indicated high viability (>85%) of the hESCs. Note that cells at the corona of the hEBs were sloughed off during the transfer process. Scale bars 500 µm.</p

hEBs (formed under -ROCK/-spin condition) expressed proteins characteristic of all the three developmental germ layers.

<p>At 20 days in suspension culture, the hEBs were positive for Alpha Fetoprotein (AFP, endoderm-specific), SOX1 (ectoderm-specific), and brachiury (mesoderm-specific) for both the (<b>a</b>) BG01V/hOG and (<b>b</b>) H9 cell lines. Cell nuclei were stained in blue with DAPI.</p

Surface sulfur content of NGFP nanofiber membrane by energy disperse.

<p>Surface sulfur content of NGFP nanofiber membrane by energy disperse.</p

Microscope observation of PC12 cells after treated with NGFP membranes containing different NGF layers for 1, 3, 5 days.

<p>NGF at a concentration of 25 ng/mL was used as the positive control. The arrows indicate neuritis associating with differentiation of PC12 cells. The scale bar is 50 μm.</p

SEM images of NGF membrane.

<p><b>PCL membrane is used as a control</b>. a) PCL membrane; b) NGFP membrane.</p

BG01V/hOG cells formed hEBs in the microwells at a cell seeding density of 15,000 hESCs per microwell.

<p>(<b>a</b>) Condensation of the hESC suspension within the microwells was progressive and evident after 6 hrs of incubation. (<b>b</b>) hEBs were compact and spherical and be able to be extracted intact from the microwells after 24 hrs of incubation. (<b>c</b>) The freshly extracted hEBs formed under four different conditions (ROCKi: ROCK inhibitor; Spin: centrifugation). (<b>d</b>) Internal structural organization among the cells within the freshly extracted hEBs (formed under -ROCK/-spin condition) was demonstrated by the presences of cell-cell junctions, i.e., adherence junctions (AJ), gap junctions (GJ), desmosomes (D), and tight junctions (TJ), in TEM images. All treatments were performed with aliquots from the same cell suspension. Microwell diameter 820 µm. Scale bars 500 µm.</p

Characteristics of TP membranes.

<p>Characteristics of TP membranes.</p

SEM images of TP membranes and their diameter distribution analysis.

<p><b>PCL membrane is used as a control</b>. a) TP7, b)TP9; c) TP11; d) PCL; e-h) size distribution of TP7, TP9 TP11 and PCL, respectively.</p

Anti-tumor effect of TP7-NGFP-TP7 device against C6 cells.

<p><b>TP7 and PCL membrane were used as controls</b>. The blank indicates the cells without treatment. <b>(*P<0.01; **P<0.001)</b>.</p