Skip to main content
. 2020 May 13;18(5):e3000705. doi: 10.1371/journal.pbio.3000705

Fig 4. The vOrganoids play important roles in the reconstruction of vascular system after transplantation.

Fig 4

(A) Schematic diagram to demonstrate the vOrganoids implantation protocols in our studies. The vOrganoids were transplanted into the S1 cortex of NOD-SCID mice. (B) Representative immunofluorescence staining figure for the classical cortical layer markers, CTIP2 and SATB2, at 2 months postimplantation. The boxed area was magnified in the right panel. Scale bar, 500 μm (left), 50 μm (right). (C) Representative immunofluorescence staining figure for presynaptic (SYB2) and postsynaptic (PSD95) in the vOrganoid grafts of 60 dpi. The displayed area was close to the graft–host border. The colocalization (arrowheads) and close association of SYB2 and PSD95 indicate that synaptic connectivity emerges in the organoid grafts. Scale bar, 10 μm. (D) The blood vessels in the vOrganoid grafts were reconstructed in three dimensions. The red tubular structure was blood vessels. Scale bar was labeled at three dimensions. Scale bar, 30 μm. (E) Immunofluorescence staining for LAMININ, dextran, and HUN were performed to confirm that the human ECs derived from vOrganoids (labeled by yellow circles and yellow arrows) and the mice ECs derived from hosts (labeled by white circles and white arrowheads) were both detected in the vascular systems in the vOrganoid grafts. Scale bar, 100 μm (left), 10 μm (right). (F) Representative immunofluorescence staining figure for GFAP and HUN in the vOrganoid grafts. A few HUN-positive astrocytes were labeled by boxes and magnified in right panels. Scale bar, 50 μm (left), 10 μm (right). (G) Representative immunofluorescence staining figure for MBP and HUN to illustrate that myelinization just could be infrequently observed at the border of graft-host commissure. The boxed areas were magnified. Scale bar, 500 μm (left), 100 μm (middle), 50 μm (right). See also S4 Fig. CTIP2, chicken ovalbumin upstream promoter transcription factor (COUP-TF)–interacting protein 2; dpi, days postimplantation; GFAP, glial fibrillary acidic protein; HUN, human nuclear; MBP, myelin basic protein; NOD-SCID, nonobese diabetic severe combined immunodeficient; PSD95, postsynaptic density protein-95; SATB2, SATB homeobox 2; SYB2, synaptobrevin 2; vOrganoid, vascularized organoid.