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Derivation and Characterization of New Human Embryonic Stem Cell Lines: SNUhES1, SNUhES2, and SNUhES3

^a Department of Obstetrics and Gynecology and
^b Institute of Reproductive Medicine and Population, Medical Research Center, College of Medicine, Seoul National University, Seoul, Korea;
^c Laboratory of Electron Microscope, Seoul National University Children’s Hospital, Seoul, Korea;
^d Department of Physiology, Yonsei University College of Medicine, Seoul, Korea

Key Words. Cell replacement therapy • Derivation of hESCs • Differentiation into cardiomyocytes • EM analysis of hESCs • Human embryonic stem cells

Correspondence: Shin Yong Moon, M.D., Ph. D., Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, 28 Yongon-dong, Chongno-gu, Seoul 110-744, Korea. Telephone: 82-2-2072-2384; Fax: 82-2-3672-7601; e-mail: shmoon{at}plaza.snu.ac.kr; and Dong-Wook Kim, Ph.D., Yonsei University College of Medicine, Department of Physiology, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-752, Korea. Telephone: 82-2-361-5208; Fax: 82-2-393-0203; e-mail: dwkim2{at}yumc.yonsei.ac.kr

Here we report the derivation and characterization of new human embryonic stem cell (hESC) lines, SNUhES1, SNUhES2, and SNUhES3. These cells, established from the inner cell mass using an STO feeder layer, satisfy the criteria that characterize pluripotent hESCs: The cell lines express high levels of alkaline phosphatase, cell surface markers (such as SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81), transcription factor Oct-4, and telomerase. When grafted into severe combined immunodeficient mice after prolonged proliferation, these cells maintained the developmental potentials to form derivatives of all three embryonic germ layers. The cell lines have normal karyotypes and distinct identities, revealed from DNA fingerprinting. Interestingly, analysis by electron microscopy clearly shows the morphological difference between undifferentiated and differentiated hESCs. Undifferentiated hESCs have a high ratio of nucleus to cytoplasm, prominent nucleoli, indistinct cell membranes, free ribosomes, and small mitochondria with a few crista, whereas differentiated cells retain irregular nuclear morphology, desmosomes, extensive cytoplasmic membranes, tonofilaments, and highly developed cellular organelles such as Golgi complex with secretory vesicles, endoplasmic reticulum studded with ribosomes, and large mitochondria. Existence of desmosomes and tonofilaments indicates that these cells differentiated into epithelial cells. When in vitro differentiation potentials of these cell lines into cardiomyocytes were examined, SNUhES3 was found to differentiate into cardiomyocytes most effectively.

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