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Open AccessArticle

Establishment of 3D Neuro-Organoids Derived from Pig Embryonic Stem-Like Cells

1
Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea
2
Institute of Stem Cell & Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju 28644, Korea
3
Institute of Animal Molecular Biotechnology, Korea University, Seoul 02841, Korea
4
Department of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea
5
Department of Neurology, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
*
Authors to whom correspondence should be addressed.
Academic Editor: Rolf Heumann
Int. J. Mol. Sci. 2021, 22(5), 2600; https://doi.org/10.3390/ijms22052600
Received: 5 February 2021 / Revised: 23 February 2021 / Accepted: 2 March 2021 / Published: 5 March 2021
Although the human brain would be an ideal model for studying human neuropathology, it is difficult to perform in vitro culture of human brain cells from genetically engineered healthy or diseased brain tissue. Therefore, a suitable model for studying the molecular mechanisms responsible for neurological diseases that can appropriately mimic the human brain is needed. Somatic cell nuclear transfer (SCNT) was performed using an established porcine Yucatan EGFP cell line and whole seeding was performed using SCNT blastocysts. Two Yucatan EGFP porcine embryonic stem-like cell (pESLC) lines were established. These pESLC lines were then used to establish an in vitro neuro-organoids. Aggregates were cultured in vitro until 61 or 102 days after neural induction, neural patterning, and neural expansion. The neuro-organoids were sampled at each step and the expression of the dopaminergic neuronal marker (TH) and mature neuronal marker (MAP2) was confirmed by reverse transcription-PCR. Expression of the neural stem cell marker (PAX6), neural precursor markers (S100 and SOX2), and early neural markers (MAP2 and Nestin) were confirmed by immunofluorescence staining. In conclusion, we successfully established neuro-organoids derived from pESLCs in vitro. This protocol can be used as a tool to develop in vitro models for drug development, patient-specific chemotherapy, and human central nervous system disease studies. View Full-Text
Keywords: porcine; neuro-organoid; embryonic stem cells; somatic cell nuclear transfer; neural differentiation porcine; neuro-organoid; embryonic stem cells; somatic cell nuclear transfer; neural differentiation
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MDPI and ACS Style

Hwang, S.-U.; Eun, K.; Kim, M.; Yoon, J.D.; Cai, L.; Choi, H.; Oh, D.; Lee, G.; Kim, H.; Kim, E.; Hyun, S.-H. Establishment of 3D Neuro-Organoids Derived from Pig Embryonic Stem-Like Cells. Int. J. Mol. Sci. 2021, 22, 2600. https://doi.org/10.3390/ijms22052600

AMA Style

Hwang S-U, Eun K, Kim M, Yoon JD, Cai L, Choi H, Oh D, Lee G, Kim H, Kim E, Hyun S-H. Establishment of 3D Neuro-Organoids Derived from Pig Embryonic Stem-Like Cells. International Journal of Molecular Sciences. 2021; 22(5):2600. https://doi.org/10.3390/ijms22052600

Chicago/Turabian Style

Hwang, Seon-Ung; Eun, Kiyoung; Kim, Mirae; Yoon, Junchul D.; Cai, Lian; Choi, Hyerin; Oh, Dongjin; Lee, Gabsang; Kim, Hyunggee; Kim, Eunhye; Hyun, Sang-Hwan. 2021. "Establishment of 3D Neuro-Organoids Derived from Pig Embryonic Stem-Like Cells" Int. J. Mol. Sci. 22, no. 5: 2600. https://doi.org/10.3390/ijms22052600

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