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

Dae Sung Kim, Dongjin R. Lee, Han Soo Kim, Jeong Eun Yoo, Sung Jun Jung, Bo Young Lim, Jiho Jang, Hoon Chul Kang, Seungkwon You, Dong Youn Hwang, Joong Woo Leem, Taick Sang Nam, Sung Rae Cho, Dong Wook Kim

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

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 hNPCPSA-NCAM+) were isolated from the heterogeneous cell population of mechanically harvested neural rosettes using magnetic-based cell sorting. The hNPCPSA-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 hNPCPSA-NCAM+ showed multipotency and responsiveness to instructive cues towards region-specific neuronal subtypes in vitro. When transplanted into the rat striatum, hNPCPSA-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 hNPCPSA-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.

Original languageEnglish
Article numbere39715
JournalPloS one
Volume7
Issue number7
DOIs
Publication statusPublished - 2012 Jul 20

Fingerprint

Neural Cell Adhesion Molecules
cell adhesion
neurons
acids
cells
Stem cells
Pluripotent Stem Cells
Cells
polysialic acid
stem cells
Nestin
Sorting
Grafts
Neurons
Rats
Screening
Contamination
Preclinical Drug Evaluations
Cell Separation
Neural Crest

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Kim, D. S., Lee, D. R., Kim, H. S., Yoo, J. E., Jung, S. J., Lim, B. Y., ... Kim, D. W. (2012). Highly pure and expandable PSA-NCAM-positive neural precursors from human ESC and iPSC-derived neural rosettes. PloS one, 7(7), [e39715]. https://doi.org/10.1371/journal.pone.0039715
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. / Highly pure and expandable PSA-NCAM-positive neural precursors from human ESC and iPSC-derived neural rosettes. In: PloS one. 2012 ; Vol. 7, No. 7.
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abstract = "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 hNPCPSA-NCAM+) were isolated from the heterogeneous cell population of mechanically harvested neural rosettes using magnetic-based cell sorting. The hNPCPSA-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 hNPCPSA-NCAM+ showed multipotency and responsiveness to instructive cues towards region-specific neuronal subtypes in vitro. When transplanted into the rat striatum, hNPCPSA-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 hNPCPSA-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.",
author = "Kim, {Dae Sung} and Lee, {Dongjin R.} and Kim, {Han Soo} and Yoo, {Jeong Eun} and Jung, {Sung Jun} and Lim, {Bo Young} and Jiho Jang and Kang, {Hoon Chul} and Seungkwon You and Hwang, {Dong Youn} and Leem, {Joong Woo} and Nam, {Taick Sang} and Cho, {Sung Rae} and Kim, {Dong Wook}",
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Kim, DS, Lee, DR, Kim, HS, Yoo, JE, Jung, SJ, Lim, BY, Jang, J, Kang, HC, You, S, Hwang, DY, Leem, JW, Nam, TS, Cho, SR & Kim, DW 2012, 'Highly pure and expandable PSA-NCAM-positive neural precursors from human ESC and iPSC-derived neural rosettes', PloS one, vol. 7, no. 7, e39715. https://doi.org/10.1371/journal.pone.0039715

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

In: PloS one, Vol. 7, No. 7, e39715, 20.07.2012.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Highly pure and expandable PSA-NCAM-positive neural precursors from human ESC and iPSC-derived neural rosettes

AU - Kim, Dae Sung

AU - Lee, Dongjin R.

AU - Kim, Han Soo

AU - Yoo, Jeong Eun

AU - Jung, Sung Jun

AU - Lim, Bo Young

AU - Jang, Jiho

AU - Kang, Hoon Chul

AU - You, Seungkwon

AU - Hwang, Dong Youn

AU - Leem, Joong Woo

AU - Nam, Taick Sang

AU - Cho, Sung Rae

AU - Kim, Dong Wook

PY - 2012/7/20

Y1 - 2012/7/20

N2 - 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 hNPCPSA-NCAM+) were isolated from the heterogeneous cell population of mechanically harvested neural rosettes using magnetic-based cell sorting. The hNPCPSA-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 hNPCPSA-NCAM+ showed multipotency and responsiveness to instructive cues towards region-specific neuronal subtypes in vitro. When transplanted into the rat striatum, hNPCPSA-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 hNPCPSA-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.

AB - 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 hNPCPSA-NCAM+) were isolated from the heterogeneous cell population of mechanically harvested neural rosettes using magnetic-based cell sorting. The hNPCPSA-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 hNPCPSA-NCAM+ showed multipotency and responsiveness to instructive cues towards region-specific neuronal subtypes in vitro. When transplanted into the rat striatum, hNPCPSA-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 hNPCPSA-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.

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