LNGFR+THY-1+ human pluripotent stem cell-derived neural crest-like cells have the potential to develop into mesenchymal stem cells

• DOI: 10.1016/j.diff.2016.04.003
• Publication Date: 2016-05-10T14:16:13Z
• Authors: Takehito Ouchi; Satoru Morikawa; Shinsuke Shibata; Kimiko Fukuda; Hironobu Okuno; Takumi Fujimura; Tatsuo Kuroda; Manabu Ohyama; Wado Akamatsu; Taneaki Nakagawa; Hideyuki Okano
• Keywords: 0301 basic medicine; Human Embryonic Stem Cells; Induced Pluripotent Stem Cells; Cell Culture Techniques; Cell Differentiation; Mesenchymal Stem Cells; Nerve Tissue Proteins; Chick Embryo; Receptors, Nerve Growth Factor; Mice; 03 medical and health sciences; Neural Crest; Animals; Humans; Thy-1 Antigens; Cell Lineage; Cell Proliferation

Mesenchymal stem cells (MSCs) are defined as non-hematopoietic, plastic-adherent, self-renewing cells that are capable of tri-lineage differentiation into bone, cartilage or fat in vitro. Thus, MSCs are promising candidates for cell-based medicine. However, classifications of MSCs have been defined retrospectively; moreover, this conventional criterion may be inaccurate due to contamination with other hematopoietic lineage cells. Human MSCs can be enriched by selection for LNGFR and THY-1, and this population may be analogous to murine PDGFRα+Sca-1+ cells, which are developmentally derived from neural crest cells (NCCs). Murine NCCs were labeled by fluorescence, which provided definitive proof of neural crest lineage, however, technical considerations prevent the use of a similar approach to determine the origin of human LNGFR+THY-1+ MSCs. To further clarify the origin of human MSCs, human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) were used in this study. Under culture conditions required for the induction of neural crest cells, human ESCs and iPSCs-derived cells highly expressed LNGFR and THY-1. These LNGFR+THY-1+ neural crest-like cells, designated as LT-NCLCs, showed a strong potential to differentiate into both mesenchymal and neural crest lineages. LT-NCLCs proliferated to form colonies and actively migrated in response to serum concentration. Furthermore, we transplanted LT-NCLCs into chick embryos, and traced their potential for survival, migration and differentiation in the host environment. These results suggest that LNGFR+THY-1+ cells identified following NCLC induction from ESCs/iPSCs shared similar potentials with multipotent MSCs.