YAP Promotes Cell Proliferation and Stemness Maintenance of Porcine Muscle Stem Cells under High-Density Condition.

Zheng Liu,Renpeng Guo,Changbo Tang,Rongrong Hu,Yichen Jiang,Ling Lin,Xi Ding,Haozhe Zhu,Zhongyuan Wu,Shijie Ding

doi:10.3390/cells10113069

Abstract

Muscle stem cells (MuSCs) isolated ex vivo are essential original cells to produce cultured meat. Currently, one of the main obstacles for cultured meat production derives from the limited capacity of large-scale amplification of MuSCs, especially under high-density culture condition. Here, we show that at higher cell densities, proliferation and differentiation capacities of porcine MuSCs are impaired. We investigate the roles of Hippo-YAP signaling, which is important regulators in response to cell contact inhibition. Interestingly, abundant but not functional YAP proteins are accumulated in MuSCs seeded at high density. When treated with lysophosphatidic acid (LPA), the activator of YAP, porcine MuSCs exhibit increased proliferation and elevated differentiation potential compared with control cells. Moreover, constitutively active YAP with deactivated phosphorylation sites, but not intact YAP, promotes cell proliferation and stemness maintenance of MuSCs. Together, we reveal a potential molecular target that enables massive MuSCs expansion for large-scale cultured meat production under high-density condition.

Highlights

Meat is a good source of high quality proteins
To determine density-mediated regulation of stemness maintenance, Muscle stem cells (MuSCs) were seeded at different densities to collagen-coated plates at 2.7 × 103/cm2 and 2.2 × 104/cm2, which were defined as low-density and high-density culture, respectively (Figure 1A)
Expression levels of MYOD and Myogenin (MYOG), two factors characterizing myogenic precursors and myogenic commitment, respectively [27], were increased to ~4.5 folds and ~9 folds in high-density cultured MuSCs compared with cells under low density condition (Figure 1D,E)

Summary

Introduction

Meat is a good source of high quality proteins. To satisfy the world’s growing population and increasing demands for food of animal origin, the meat industry would have to increase production by approximately 50–100% in the three decades [1]. It is necessary to develop new technologies for sustainable meat production and supply. Lab-based cell cultured meat production is achieved through isolation and expansion of animal stem cells, committed myogenic or adipogenic differentiation, and food processing [3]. In contrast to traditional meat production, cultured meat technology exhibits distinct advantages in sustainability, animal welfare and public health [4]. Cultured meat has been tasted in the UK, China and other parts of the world. In 2013, the first cell-cultured hamburger, which was developed by Dr Mark Post at the University of Maastricht, was cooked and tasted in London, England [5]. In China, cultured meat was firstly tasted in Nanjing Agricultural University in 2020, which was pioneered by Dr Guanghong Zhou

Methods

Results

Conclusion