Baek Jin Ah, Seol Hye Won, Jung Juwon, Kim Hee Sun, Oh Sun Kyung, Choi Young Min
1Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, 71, Ihwajang-gil, Jongno-gu, Seoul, 03087 Korea.
2Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, 101, Daehak-ro, Jongno-gu, Seoul, 03080 Korea.
Tissue Eng Regen Med. 2017 Jun 8;14(4):453-464. doi: 10.1007/s13770-017-0053-2. eCollection 2017 Aug.
Human embryonic stem cell (hESC) culture system has been changing culture conditions from conventional to xeno-free for therapeutic cell applications, and -glycolylneuraminic acid (Neu5Gc) could be a useful indicator of xenogeneic contaminations in hESCs because human cells can no longer produce it genetically. We set up the humanized culture condition using commercially available humanized materials and two different adaptation methods: sequential or direct. SNUhES4 and H1 hESC lines, previously established in conventional culture conditions, were maintained using the humanized culture condition and were examined for the presence of Neu5Gc. The hESCs showed the same morphology and character as those of the conventional culture condition. Moreover, they were negative for Neu5Gc within two passages without loss of pluripotency. This study suggested that this method can effectively cleanse previously established hESC lines, bringing them one step closer to being clinical-grade hESCs.
人类胚胎干细胞（hESC）培养系统一直在改变培养条件，从传统培养转变为无动物源培养以用于治疗性细胞应用，而N-羟乙酰神经氨酸（Neu5Gc）可能是hESC中异种污染的一个有用指标，因为人类细胞在基因层面上已无法再产生它。我们使用市售的人源化材料和两种不同的适应方法（顺序适应法或直接适应法）建立了人源化培养条件。先前在传统培养条件下建立的SNUhES4和H1 hESC系，采用人源化培养条件进行维持，并检测Neu5Gc的存在情况。这些hESC表现出与传统培养条件下相同的形态和特性。此外，它们在传代两次内Neu5Gc呈阴性，且未丧失多能性。这项研究表明，该方法可以有效地清除先前建立的hESC系，使其更接近临床级hESC。
