Highly pure and expandable PSA-NCAM-positive neural precursors from human ESC and iPSC-derived neural rosettes

• Published in: PloS one Vol. 7; no. 7; p. e39715
• Main Authors: Kim, Dae-Sung, Lee, Dongjin R, Kim, Han-Soo, Yoo, Jeong-Eun, Jung, Sung Jun, Lim, Bo Young, Jang, Jiho, Kang, Hoon-Chul, You, Seungkwon, Hwang, Dong-Youn, Leem, Joong Woo, Nam, Taick Sang, Cho, Sung-Rae, Kim, Dong-Wook
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 20-07-2012; Public Library of Science (PLoS)
• Subjects: Adhesive strength; Animals; Astrocytes; Biology; Biomarkers; Biomedical materials; Brain - cytology; Brain research; Cell adhesion; Cell adhesion molecules; Cell culture; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell surface; Cell Survival; Cells, Cultured; Contamination; Cues; Cultivation; Drug screening; Embryos; Epidermal growth factor; Female; Gene expression; Grafts; Humans; Immunomagnetic Separation; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Induced Pluripotent Stem Cells - transplantation; Intermediate filament proteins; Medicine; Musashi protein; Neostriatum; Nervous system; Nervous system diseases; Nestin; Neural cell adhesion molecule; Neural Cell Adhesion Molecules - metabolism; Neural crest; Neural stem cells; Neural Stem Cells - cytology; Neural Stem Cells - metabolism; Neural Stem Cells - transplantation; Neurodegenerative diseases; Neurons; Neurons - cytology; Neurons - metabolism; Oligodendrocytes; Physicians; Physiology; Pluripotency; Polysialic acid; Precursors; Rats; Rats, Sprague-Dawley; Sialic Acids - metabolism; Spinal cord; Stem Cell Transplantation; Stem cells; Studies; Surface markers; Telomerase; Tumorigenesis
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0039715

Homogeneous culture of neural precursor cells (NPCs) derived from human pluripotent stem cells (hPSCs) would provide a powerful tool for biomedical applications. However, previous efforts to expand mechanically dissected neural rosettes for cultivation of NPCs remain concerns regarding non-neural cell contamination. In addition, several attempts to purify NPCs using cell surface markers have not demonstrated the expansion capability of the sorted cells. In the present study, we show that polysialic acid-neural cell adhesion molecule (PSA-NCAM) is detected in neural rosette cells derived from hPSCs, and employ PSA-NCAM as a marker for purifying expandable primitive NPCs from the neural rosettes. PSA-NCAM-positive NPCs (termed hNPC(PSA-NCAM+)) were isolated from the heterogeneous cell population of mechanically harvested neural rosettes using magnetic-based cell sorting. The hNPC(PSA-NCAM+) extensively expressed neural markers such as Sox1, Sox2, Nestin, and Musashi-1 (80∼98% of the total cells) and were propagated for multiple passages while retaining their primitive characteristics in our culture condition. Interestingly, PSA-NCAM-negative cells largely exhibited characteristics of neural crest cells. The hNPC(PSA-NCAM+) showed multipotency and responsiveness to instructive cues towards region-specific neuronal subtypes in vitro. When transplanted into the rat striatum, hNPC(PSA-NCAM+) differentiated into neurons, astrocytes, and oligodendrocytes without particular signs of tumorigenesis. Furthermore, Ki67-positive proliferating cells and non-neural lineage cells were rarely detected in the grafts of hNPC(PSA-NCAM+) compared to those of neural rosette cells. Our results suggest that PSA-NCAM-mediated cell isolation provides a highly expandable population of pure primitive NPCs from hPSCs that will lend themselves as a promising strategy for drug screening and cell therapy for neurodegenerative disorders.