Biomaterials Translational ›› 2024, Vol. 5 ›› Issue (1): 21-32.doi: 10.12336/biomatertransl.2024.01.003
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Zhangjie Li1, Dingyuan Yu1, Chenyang Zhou1, Feifan Wang1, Kangyi Lu1, Yijun Liu1, Jiaqi Xu1, Lian Xuan2, Xiaolin Wang1,2,3,4,*()
Received:
2024-01-16
Revised:
2024-02-29
Accepted:
2024-03-14
Online:
2024-03-28
Published:
2024-03-28
Contact:
Xiaolin Wang, About author:
Xiaolin Wang, xlwang83@sjtu.edu.cn.Figure 1. Schematic representation of the principles underlying two methods and top-down EC lining method for constructing vascularised models in vitro. (A) Schematic illustrating two methods for constructing vascularised models in vitro: (i) Principle of the endothelial cell lining method. (ii) Principle of the self-assembly method. Created with BioRender.com. (B) Microvascular system combined with viscous finger patterning technique: (i) Schematic of the microfluidic chip showing dimensions and layout of the microfluidic platform, four straight channels on a single chip with designed parameters 500 μm × 500 μm × 1 cm (w × h × l). (ii) 20× magnification confocal slice of a patterned lumen with 5 mg/mL collagen I. Scale bar: 200 μm. (iii) XZ reconstruction showing the flow field of the scaffold. Scale bar: 200 μm. Reprinted from de Graaf et al.38 (C) Spiral microvascular system: (i) Cross-sectional fluorescence image of the spiral blood vessel. (ii) Spiral vascularised tumour model demonstrating vascular growth towards tumour cells. (iii) Spiral vascularised heart model. Scale bars: 200 μm. Reprinted from Mandrycky et al.24 CM: cardiomyocyte; cTnT: cardiac troponin T; EC: endothelial cell; hESC: human embryonic stem cell.
Figure 2. Bottom-up self-assembly vascularisation method for constructing vascularised models in vitro. (A) The modular microfluidic system combines two PDMS layers: (i) Medium channel module (upper layer). (ii) Tissue chamber module (bottom layer). (iii) A completed two-layered device shows the medium channels and tissue chambers in different layers. (iv) Large-scale perfused microvascular networks are generated using different configurations. Reprinted from Yue et al.50 (B) Design diagram and overview of the microfluidic system with perfusion vascular network: (i) Photograph of a microfluidic device. (ii) Schematic diagram showing an overview of assays. Reprinted from Nashimoto et al. 51 Copyright 2017 The Royal Society of Chemistry. Reproduced with permission. (C) Design diagram and representative images of the microfluidic vascular model as a non-contact cell culture device: (i) Photograph of the microfluidic chip used in the study to generate a perfusable vascular network. (ii) Representative images show the comparison of cocultured ECs to monocultured ECs over 3 days. Scale bars: 100 μm. Reprinted from Tan et al.53 EC: endothelial cell; FB: fibroblast: PDMS: polydimethylsiloxane.
Figure 3. Hybrid method for constructing vascularised models in vitro. (A) Vascularised model construction combining vasculogenesis mechanism and EC lining method: (i) Schematic diagram of the microfluidic chip. The physiological level of mechanical stimulion vasculogenesis was established by filling the reservoirs to different culture medium heights (V1: 23 mm H2O, V2: 8 mm H2O, V3: 18 mm H2O, V4: 3 mm H2O (1 mm H2O = 9.80665 Pa)). (ii) Microchannel design. The gel loading port (a, b), the media reservoir ports (c-f), cell loading port (g-j). (iii) Observation of angiogenesis phenomenon in the microfluidic chip. Scale bars: 100 μm. GFP (green)-labeled indicates the lined ECs at the cell-matrix interface invaded into the 3D fibrin gel and formed microvascular sprouts as early as 24 hours post-lining. From inside the tissue chamber, mCherry-expressing (red) ECs (ECs-mCherry) migrated outward to the laminin-coated microfluidic channel to connect with the lined BFP-labeled (blue) ECs (ECs-BFP) in the channels. The green image below represents more time of microvascular sprouts invasion than the green image above. Reprinted from Wang et al.54 Copyright 2016 The Royal Society of Chemistry. (B) Diagram of the impact of chip implantation on maternal vascular remodelling: (i) Interactions between invading EVTs and maternal blood vessels were modeled in the implantation-on-a-chip. (ii) 3D projection images of EVTs (green) migrating across the ECM hydrogel towards maternal ECs. Scale bar: 200 μm. Reprinted from Park et al.55 (C) MSVT construction: (i) Schematic of the MSVT fabrication process. (ii) Maximum intensity projection of the MSVT, with high magnifications. The middle image is an enlargement of the dashed box in the left image. The right image is an enlargement of the dashed box in the middle image. The white arrows of the right image indicate micro-vessels sprouts and the dashed lines of the right image indicate the macro-vessels borders. Scale bars: 500 μm (left), 100 μm (middle), 50 μm (right). Reprinted from Debbi et al.56 (D) Fluorescent bead perfusion of engineered microvessels (μVs) and citrated whole-blood perfusion in μVs: (i) Brightfield stitched large image of red blood cell-filled pattern and sprouts with magnified view (inset, white dotted boundary) for hESC-ECs-seeded μV only constructs after 4 days of culture. Scale bar: 200 μm. (ii) μV + SA construct with perfused anastomotic connections. High magnification views of outlined regions 1 and 2 with corresponding in situ staining for mTm-hESC-ECs (DsRed+, red) and GFP-hESC-ECs (GFP+, green). Scale bars: 100 μm (left), 40 μm (right). Reprinted from Redd et al.57 BFP: blue fluorescent protein; EC: endothelial cell; ECM: extracellular matrix; EVT: extravillous trophoblast; GFP: green fluorescent protein; hESC-EC: human embryonic stem cell-derived endothelial cell; MSVT: multi-scale vascular tissue; SA: self-assembled.
Study | Method | Cells used | Minimum diameter | Vascular network shape |
---|---|---|---|---|
Qiu et al. | Top-down EC lining method | HUVECs, HDMVECs, HLMVECs | ~20 μm | Cylindrical |
de Graaf et al. | Top-down EC lining method | iPSC-ECs, HBVPs | ~300 μm | Cylindrical |
Mandrycky et al. | Top-down EC lining method | HUVECs | ~200 μm | Spiral |
Miller et al. | Top-down EC lining method | HUVECs, 10T1/2 cells | ~300 μm | Cylindrical |
Yue et al. | Bottom-up self-assembly vascularisation method | ECFC-ECs, NHLFs | ~10 μm | Capillary |
Nashimoto et al. | Bottom-up self-assembly vascularisation method | HUVECs, NHLFs | ~10 μm | Capillary |
Phan et al. | Bottom-up self-assembly vascularisation method | ECFC-ECs, NHLFs | ~20 μm | Capillary |
Tan et al. | Bottom-up self-assembly vascularisation method | HUVECs, NHLFs | ~20 μm | Capillary |
Wang et al. | Hybrid method | ECFC-ECs, NHLFs | ~15 μm | Multi-scale vascular network |
Park et al. | Hybrid method | HEMVECs, HLMVECs, HBMVECs, NHLFs | ~10 μm | Multi-scale vascular network |
Debbi et al. | Hybrid method | HAMEC, DPSCs | ~20 μm | Multi-scale vascular network |
Rebbi et al. | Hybrid method | hESC-ECs, HUVECs | ~20 μm | Multi-scale vascular network |
Table 1. Comparison of different in vitro vascularisation model construction methods
Study | Method | Cells used | Minimum diameter | Vascular network shape |
---|---|---|---|---|
Qiu et al. | Top-down EC lining method | HUVECs, HDMVECs, HLMVECs | ~20 μm | Cylindrical |
de Graaf et al. | Top-down EC lining method | iPSC-ECs, HBVPs | ~300 μm | Cylindrical |
Mandrycky et al. | Top-down EC lining method | HUVECs | ~200 μm | Spiral |
Miller et al. | Top-down EC lining method | HUVECs, 10T1/2 cells | ~300 μm | Cylindrical |
Yue et al. | Bottom-up self-assembly vascularisation method | ECFC-ECs, NHLFs | ~10 μm | Capillary |
Nashimoto et al. | Bottom-up self-assembly vascularisation method | HUVECs, NHLFs | ~10 μm | Capillary |
Phan et al. | Bottom-up self-assembly vascularisation method | ECFC-ECs, NHLFs | ~20 μm | Capillary |
Tan et al. | Bottom-up self-assembly vascularisation method | HUVECs, NHLFs | ~20 μm | Capillary |
Wang et al. | Hybrid method | ECFC-ECs, NHLFs | ~15 μm | Multi-scale vascular network |
Park et al. | Hybrid method | HEMVECs, HLMVECs, HBMVECs, NHLFs | ~10 μm | Multi-scale vascular network |
Debbi et al. | Hybrid method | HAMEC, DPSCs | ~20 μm | Multi-scale vascular network |
Rebbi et al. | Hybrid method | hESC-ECs, HUVECs | ~20 μm | Multi-scale vascular network |
Figure 4. Strategies for organoid vascularisation. (A) Schematic representation of the principles underlying three strategies. Created with BioRender.com. (B) Representative immunofluorescence staining figure for CTIP2/IB4/PAX6 at day 65 to demonstrate that the vascular (IB4) structures would progressively extend into newborn neurons (CTIP2) with the development of vOrganoids. Scale bar: 50 μm. Reprinted from Shi et al.59 (C) Immunostaining of whole-mount vhCOs and control hCOs at different time points (days 30 and 70) for DAPI/CD31/MAP2. It illustrates that vhCOs had significantly more vessel area and length than control hCOs. Scale bars: 100 μm. Reprinted from Cakir et al.62 (D) Whole-mount staining results in the fused vasculature and brain organoids. D40-fVBOr represents the fused vasculature and brain organoids at day 40. Reprinted from Sun et al.65 Scale bar: 200 μm. CTIP2: COUP TF1 interacting protein 2; DAPI: 4′,6-diamidino-2-phenylindole; DCX: doublecortin; EC: endothelial cell; GFP: green fluorescent protein; hCOs: human cortical organoids; IB4: isolectin B4; MAP2: microtubule-associated protein 2; PAX6: paired box 6; PSC: pluripotent stem cell; vhCOs: vascularised human cortical organoids; vOrganoids: vascularised organoids.
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