A simple method to improve the quality and yield of human pluripotent stem cell-derived cerebral organoids

Mu Seog Choe; So Jin Kim; Seung Taek Oh; Chang Min Bae; Won-Young Choi; Kyung Min Baek; Joong Sun Kim; Min Young Lee

doi:10.1016/j.heliyon.2021.e07350

A simple method to improve the quality and yield of human pluripotent stem cell-derived cerebral organoids

Heliyon. 2021 Jun 19;7(6):e07350. doi: 10.1016/j.heliyon.2021.e07350. eCollection 2021 Jun.

Authors

Mu Seog Choe^{1

2}, So Jin Kim^{1

2}, Seung Taek Oh^{1

2}, Chang Min Bae^{1

2}, Won-Young Choi^{1

2}, Kyung Min Baek³, Joong Sun Kim⁴, Min Young Lee^{1

2}

Affiliations

¹ College of Pharmacy, Research Institute of Pharmaceutical Sciences Kyungpook National University, Daegu, South Korea.
² College of Pharmacy, Pharmaceutical Sciences, Vessel-Organ Interaction Research Center (VOICE, MRC), Kyungpook National University, Daegu, South Korea.
³ Department of Cardiovascular and Neurologic Disease, College of Oriental Medicine, Daegu Haany University, Daegu, South Korea.
⁴ K-herb Research Center, Korea Institute of Oriental Medicine, Daejeon, South Korea.

Abstract

The development of cerebral organoid technology has allowed the human neural tissue to be collected for studying human brain development and neurological diseases. Human pluripotent stem cell-derived cerebral organoids (hCOs) are a theoretically infinite source of fresh human brain tissue for various research purposes. However, hCOs have limitations, including core necrotic cell death. To solve this problem, we tested a simple method, which has been previously overlooked. In this study, we mechanically cut 70-day-old hCOs with a scalpel blade into 2 to 4 pieces, each depending on their original size. After culturing cut hCOs for additional 7 days, their size was less variable and smaller than uncut hCOs and there were no histological differences between uncut and cut hCOs. Note that hypoxia-inducible factor (HIF)-1α was expressed in the central area of uncut hCOs but not in cut hCOs. Uncut hCOs, therefore, showed broad core areas stained with terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL), whereas cut hCOs did not. In conclusion, this simple mechanical cutting method allowed us to acquire a larger number of hCOs without a necrotic core.

Keywords: Cerebral organoids; Human pluripotent stem cell; Mechanical cutting; Necrosis.