Construction of transplantable artificial vascular tissue based on adipose tissue-derived mesenchymal stromal cells by a cell coating and cryopreservation technique

Yoshiya Asano,Tetsuro Watabe,Mitsuru Akashi,Yasuhiro Yoshimatsu,Daisuke Okano,Michiya Matsusaki,Hiroshi Shimoda

doi:10.1038/s41598-021-97547-2

Abstract

Prevascularized artificial three-dimensional (3D) tissues are effective biomaterials for regenerative medicine. We have previously established a scaffold-free 3D artificial vascular tissue from normal human dermal fibroblasts (NHDFs) and umbilical vein-derived endothelial cells (HUVECs) by layer-by-layer cell coating technique. In this study, we constructed an artificial vascular tissue constructed by human adipose tissue-derived stromal cells (hASCs) and HUVECs (ASCVT) by a modified technique with cryopreservation. ASCVT showed a higher thickness with more dense vascular networks than the 3D tissue based on NHDFs. Correspondingly, 3D-cultured ASCs showed higher expression of several angiogenesis-related factors, including vascular endothelial growth factor-A and hepatic growth factor, compared to that of NHDFs. Moreover, perivascular cells in ASCVT were detected by pericyte markers, suggesting the differentiation of hASCs into pericyte-like cells. Subcutaneous transplantation of ASCVTs to nude mice resulted in an engraftment with anastomosis of host’s vascular structures at 2 weeks after operation. In the engrafted tissue, the vascular network was surrounded by mural-like structure-forming hASCs, in which some parts developed to form vein-like structures at 4 weeks, suggesting the generation of functional vessel networks. These results demonstrated that cryopreserved human cells, including hASCs, could be used directly to construct the artificial transplantable tissue for regenerative medicine.

Highlights

Prevascularized artificial three-dimensional (3D) tissues are effective biomaterials for regenerative medicine
Vascular network formation in human adipose tissue-derived stromal cells (hASCs)‐based 3D artificial vascular tissue fabricated by Cryopreserved cell accumulation method (CP‐CAM)
We firstly examined the vessel network formation using artificial vascular tissue constructed by normal human dermal fibroblasts (NHDFs) and HUVECs, termed as FbVT (Supplementary information, Fig. S1–S4)

Summary

Introduction

Prevascularized artificial three-dimensional (3D) tissues are effective biomaterials for regenerative medicine. For fabrication of vascular tissue, the fibronectin and gelatin-coated endothelial cells are cultivated between the layers of normal human dermal fibroblasts (NHDFs) They rapidly form a network of blood and/or lymphatic vessel-like tubules within a week[9,10]. The lymphatic vascular tissue constructed using NHDFs and human dermal lymphatic endothelial cells showed remodeling of the network with generation of blood vessel-like structures after engraftment into the back fascia or subcutaneous tissue of nude m ice[4] From these studies, we proposed the potency of the artificial vascular tissues constructed by the cell accumulation method in regenerative m edicine[3,4]

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