Printability, Durability, Contractility and Vascular Network Formation in 3D Bioprinted Cardiac Endothelial Cells Using Alginate–Gelatin Hydrogels

The blood islands of the visceral yolk sac (VYS) are the initial sites of hematopoiesis in mammals. We have developed a yolk sac explant culture system to study the process of blood cell and endothelial cell development from extraembryonic mesoderm cells. No benzidine-positive cells or beta H1-globin mRNA expression was detected at the primitive streak or neural plate stage of development (E7.5). However, when isolated E7.5 dissected tissues were cultured for 36 to 72 hours in serum-free medium, hundreds of hemoglobin-producing cells and embryonic globin gene expression were identified in both intact yolk sac and VYS mesoderm explants. Explanted E7.5 extraembryonic mesoderm tissues thus recapitulate in vivo primitive erythropoiesis and do not require the presence of a vascular network or the VYS endoderm. Yolk sac blood islands also contain endothelial cells that arise by vasculogenesis and express flk-1. We detected flk-1 mRNA as early as the primitive streak stage of mouse embryogenesis. Culture of embryo proper and intact VYS explants, which contain both mesoderm and endoderm cells, produced capillary networks and expressed flk-1. In contrast, vascular networks were not seen when VYS mesoderm was cultured alone, although flk-1 expression was similar to that of intact VYS explants. The addition of vascular endothelial growth factor to VYS mesoderm explants did not induce vascular network formation. These results suggest that the VYS endoderm or its extracellular matrix is necessary for the coalescence of developing endothelial cells into capillary networks.

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Vascular network formation for a long-term spheroid culture by co-culturing endothelial cells and fibroblasts

2015 28th IEEE International Conference on Micro Electro Mechanical Systems (MEMS) ◽

10.1109/memsys.2015.7050995 ◽

2015 ◽

Cited By ~ 4

Author(s):

Tomoya Hayashi ◽

Hisako Takigawa-Imamura ◽

Koichi Nishiyama ◽

Hirofumi Shintaku ◽

Hidetoshi Kotera ◽

...

Keyword(s):

Endothelial Cells ◽

Network Formation ◽

Vascular Network ◽

Spheroid Culture

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Endothelial cell crosstalk improves browning but hinders white adipocyte maturation in 3D engineered adipose tissue

Integrative Biology ◽

10.1093/intbio/zyaa006 ◽

2020 ◽

Vol 12 (4) ◽

pp. 81-89

Author(s):

Jennifer H Hammel ◽

Evangelia Bellas

Keyword(s):

Endothelial Cells ◽

Adipose Tissue ◽

Type I Collagen ◽

Network Formation ◽

Uncoupling Protein ◽

Collagen Gel ◽

Cell Types ◽

Vascular Network ◽

Type I ◽

White Adipocyte

Abstract Central to the development of adipose tissue (AT) engineered models is the supporting vasculature. It is a key part of AT function and long-term maintenance, but the crosstalk between adipocytes and endothelial cells is not well understood. Here, we directly co-culture the two cell types at varying ratios in a 3D Type I collagen gel. Constructs were evaluated for adipocyte maturation and function and vascular network organization. Further, these constructs were treated with forskolin, a beta-adrenergic agonist, to stimulate lipolysis and browning. Adipocytes in co-cultures were found to be less mature than an adipocyte-only control, shown by smaller lipid droplets and downregulation of key adipocyte-related genes. The most extensive vascular network formation was found in the 1:1 co-culture, supported by vascular endothelial growth factor (VEGF) upregulation. After forskolin treatment, the presence of endothelial cells was shown to upregulate PPAR coactivator 1 alpha (PGC-1α) and leptin, but not uncoupling protein 1 (UCP1), suggesting a specific crosstalk that enhances early stages of browning.

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Vascular Network Formation by Human Microvascular Endothelial Cells in Modular Fibrin Microtissues

ACS Biomaterials Science & Engineering ◽

10.1021/acsbiomaterials.6b00274 ◽

2016 ◽

Vol 2 (11) ◽

pp. 1914-1925 ◽

Cited By ~ 19

Author(s):

Ramkumar Tiruvannamalai Annamalai ◽

Ana Y. Rioja ◽

Andrew J. Putnam ◽

Jan P. Stegemann

Keyword(s):

Endothelial Cells ◽

Network Formation ◽

Vascular Network ◽

Microvascular Endothelial Cells ◽

Microvascular Endothelial

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Spatial patterning of endothelial cells and vascular network formation using ultrasound standing wave fields

The Journal of the Acoustical Society of America ◽

10.1121/1.4812867 ◽

2013 ◽

Vol 134 (2) ◽

pp. 1483-1490 ◽

Cited By ~ 20

Author(s):

Kelley A. Garvin ◽

Diane Dalecki ◽

Mohammed Yousefhussien ◽

Maria Helguera ◽

Denise C. Hocking

Keyword(s):

Endothelial Cells ◽

Standing Wave ◽

Network Formation ◽

Vascular Network ◽

Spatial Patterning ◽

Wave Fields

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Stromal cells expressing ephrin-B2 promote the growth and sprouting of ephrin-B2+ endothelial cells

Blood ◽

10.1182/blood.v98.4.1028 ◽

2001 ◽

Vol 98 (4) ◽

pp. 1028-1037 ◽

Cited By ~ 52

Author(s):

Xiu-Qin Zhang ◽

Nobuyuki Takakura ◽

Yuichi Oike ◽

Tomohisa Inada ◽

Nicholas W. Gale ◽

...

Keyword(s):

Endothelial Cells ◽

Stromal Cells ◽

Network Formation ◽

Smooth Muscle Actin ◽

Cell System ◽

Vascular Network ◽

Functional Studies ◽

Vasculogenesis And Angiogenesis ◽

Arterial Endothelial Cells

Ephrin-B2 is a transmembrane ligand that is specifically expressed on arterial endothelial cells (ECs) and surrounding cells and interacts with multiple EphB class receptors. Conversely, EphB4, a specific receptor for ephrin-B2, is expressed on venous ECs, and both ephrin-B2 and EphB4 play essential roles in vascular development. The bidirectional signals between EphB4 and ephrin-B2 are thought to be specific for the interaction between arteries and veins and to regulate cell mixing and the making of particular boundaries. However, the molecular mechanism during vasculogenesis and angiogenesis remains unclear. Manipulative functional studies were performed on these proteins in an endothelial cell system. Using in vitro stromal cells (OP9 cells) and a paraaortic splanchnopleura (P-Sp) coculture system, these studies found that the stromal cells expressing ephrin-B2 promoted vascular network formation and ephrin-B2+ EC proliferation and that they also induced the recruitment and proliferation of α-smooth muscle actin (α-SMA)–positive cells. Stromal cells expressing EphB4 inhibited vascular network formation, ephrin-B2+ EC proliferation, and α-SMA+ cell recruitment and proliferation. Thus, these data suggest that ephrin-B2 and EphB4 mediate reciprocal interactions between arterial and venous ECs and surrounding cells to form each characteristic vessel.

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