Human iPSC‐Vascular smooth muscle cell spheroids demonstrate size‐dependent alterations in cellular viability and secretory function

Abstract

Human-induced pluripotent stem cells (hiPSC) and their differentiated vascular cells have been revolutionizing the field of regenerative wound healing. These cells are shown to be rejuvenated with immense potentials in secreting paracrine factors.Recently, hiPSC-derived vascular smooth muscle cells (hiPSC-VSMC) have shown regenerative wound healing ability via their paracrine secretion. The quest to modulate the secretory function of these hiPSC-VSMC is an ongoing effort and involves the use of both biochemical and biophysical stimuli. This study explores the development and optimization of a reproducible, inexpensive protocol to form hiPSC-VSMC derived spheroids to investigate the implications of spheroid size on viability and paracrine secretion. Our data show the successful formation of different sizes of spheroids using various amount of hiPSC-VSMC. The hiPSC-VSMC spheroids formed with 10,000 cells strike an ideal balance between overall cell health and maximal paracrine secretion. The conditioned medium from these spheroids was found to be bioactive in enhancing human dermal fibroblast cell proliferation and migration. This research will inform future studies on the optimal spheroid size for regenerative wound healing applications.

Use of iPSC-Derived Smooth Muscle Cells to Model Physiology and Pathology

“…91,92 Some of the most common and recently developed 3D cell culture techniques for hiPSC-derived SMCs involve the usage of organ-on-a-chip, vascular cell spheroids, and engineered vascular tissues (eVTs) to name a few. [93][94][95] The ongoing diversification of these techniques has exponentially increased within the last decade, attracting more attention to the potential that 3D models have for the study of vascular function and disease. Threedimensional cultures allow for the proper formation of and interaction with the ECM, mimicking cell adhesion behaviors as well as migratory and differentiation cues that are present in vivo.…”

“…98,99 Table 1 comprises a list of some of the most recently published works that integrate the usage of tissue engineering methods for hiPSC-SMCs, their application, and their relevance. 51,94,[100][101][102][103][104][105][106][107][108] Future Clinical Uses of hiPSC-SMCs Transplantation of vascular cells to revascularize damaged tissue and improve tissue perfusion is a potential therapeutic strategy for many diseases. 109,110 Cotransplantation of hiPSC-SMCs with other iPSC-derived cells such as ECs has been shown to efficiently mediate key processes such as neovascularization.…”

“…98,99 Table 1 comprises a list of some of the most recently published works that integrate the usage of tissue engineering methods for hiPSC-SMCs, their application, and their relevance. 51,94,100–108…”

Induced pluripotent stem cells for cardiovascular therapeutics: Progress and perspectives

“…VSMCs derived from human iPSCs can also provide an unlimited cell source for autologous cell-based therapies and vascular disease models, as well as for studying pathophysiological mechanisms (Table 1). Differentiated iPSC-derived VSMCs express myogenic proteins such as ACTA2, TAGLN, MYH10, CALM1, CNN1, CALD1, myosin light chain genes, MYLK1, and late VSMCs differentiation marker MYH11, and demonstrate fibrillar organization and contractile responses to receptor-and non-receptor-mediated agonists characteristic of the development of VSMCs with the contractile phenotype (Bajpai et al, 2012;Biel et al, 2015;Patsch et al, 2015;Paik et al, 2018;Kwong et al, 2019;Duan et al, 2021;Islam et al, 2022). Cell surface markers such as PDGFRB (CD140b) and LRP1 (CD91) are present in both iPSC-derived progenitor and mature VSMCs (Biel et al, 2015).…”