Spatially Assembled Bilayer Cell Sheets of Stem Cells and Endothelial Cells Using Thermosensitive Hydrogels for Therapeutic Angiogenesis

Abstract

Although the coculture of multiple cell types has been widely employed in regenerative medicine, in vivo transplantation of cocultured cells while maintaining the hierarchical structure remains challenging. Here, a spatially assembled bilayer cell sheet of human mesenchymal stem cells and human umbilical vein endothelial cells on a thermally expandable hydrogel containing fibronectin is prepared and its effect on in vitro proangiogenic functions and in vivo ischemic injury is investigated. The expansion of hydrogels in response to a temperature change from 37 to 4 °C allows rapid harvest and delivery of the bilayer cell sheet to two different targets (an in vitro model glass surface and in vivo tissue). The in vitro study confirms that the bilayer sheet significantly increases proangiogenic functions such as the release of nitric oxide and expression of vascular endothelial cell genes. In addition, transplantation of the cell sheet from the hydrogels into a hindlimb ischemia mice model demonstrates significant retardation of necrosis particularly in the group transplated with the bilayer sheet. Collectively, the bilayer cell sheet is readily transferrable from the thermally expandable hydrogel and represents an alternative approach for recovery from ischemic injury, potentially via improved cell-cell communication.

Fibronectin-Enriched Biomaterials, Biofunctionalization, and Proactivity: A Review

“…Hydrogels could be obtained with different types of hydrophilic polymers (natural or synthetic). However, there may be some drawbacks, such as insufficient cell-anchorage cues, which can lead to minimal cell adhesion and spreading [99][100][101] or a decrease in cell viability in the case of cell encapsulation [102,103].…”

“…Some remarkable studies dealing with Fn physical dispersion have been reported for alginate-based hydrogels [98,100,101,113]. For instance, Mosahebi et al investigated the addition of Fn in an alginate solution before physical gelation with Ca 2+ and its impact on nerve regeneration.…”

Advances in the Application of Biomimetic Endometrium Interfaces for Uterine Bioengineering in Female Infertility

“…Both of them contained abundant extracellular matrix (ECM) necessary for providing maximum strength and adherence to the host tissue surface . CSE includes cell culture in thermo-sensitive plates that develop compact cell sheets which detach with temperature drop (Jun et al, 2017). As a major advantage, this technology permits maintenance of growth factors, ECM proteins, and other bioactive cytokines because cells are detached spontaneously from the culture plates without the need for enzymatic procedures and proteolytic treatments.…”

Design of Polymeric Culture Substrates to Promote Proangiogenic Potential of Stem Cells

“…This approach significantly increased proangiogenic activities by stimulating the release of nitric oxide and the expression of various types of proangiogenic factors (VE‐cadherin, vWF, and Tie‐2). Moreover, necrosis was significantly retarded after transplantation of cells into a region of hindlimb ischemia in mice, particularly in the group treated with the bilayer sheet . In another example, a human platelet lysate (PL) hydrogel was applied as a scaffold for delivery of MSCs and HUVECs to promote vascular regeneration.…”