Human primary cultured hepatic stellate cells can be cryopreserved (original) (raw)

Abstract

We compared the morphological and functional characteristics of cultured unfrozen hepatic stellate cells (HSCs) and cryopreserved HSCs obtained from human livers. We used liver tissues obtained by surgical resection from patients with metastatic liver cancer or with hepatocellular carcinoma. HSCs were isolated and allowed to spread in culture. Comparison of morphological and functional features between the unfrozen HSCs and cryopreserved HSCs was performed at each passage using the following techniques: light microscopy, immunohistochemistry, cell growth curve, metallothionein (MTT) assay, and PI staining, Western blot, real-time polymerase chain reaction (PCR), and gene expression analysis using microarrays. The purity of HSCs was more than 90% in all passages. α-Smooth muscle actin (SMA-)positive HSCs gradually increased in successive passages, and the positive cell rate and rate of increase in cell number were similar in both groups. Expression of platelet-derived growth factor (PDGF) receptor, transforming growth factor (TGF)-β receptor, and α-SMA mRNAs and protein was similar during each passage in the two groups. Gene expression was nearly identical at each passage in unfrozen and frozen/thawed samples obtained from the same patient. In conclusion, an adequate protocol for the cryopreservation of human primary cultured HSCs could be established.

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Authors and Affiliations

  1. Research Center for Innovative Cancer Therapy, Kurume University, 67 Asahi-machi, Kurume, 830-0011, Japan
    Anna Nakamura, Takato Ueno, Yumihiko Yagi, Koji Okuda, Toshiro Ogata, Toru Nakamura, Takuji Torimura, Hideki Iwamoto, Sivakumar Ramadoss & Michio Sata
  2. Department of Experimental Pathology, Center for Research and Advanced Studies, National Polytechnic Institute, Mexico City, Mexico
    Victor Tsutsumi
  3. Cell-Innovator, Venture Business Laboratory, Kyushu University, Fukuoka, Japan
    Kaori Yasuda & Yumi Tomiyasu
  4. Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan
    Kenichi Obayashi, Kosuke Tashiro & Satoru Kuhara

Authors

  1. Anna Nakamura
  2. Takato Ueno
  3. Yumihiko Yagi
  4. Koji Okuda
  5. Toshiro Ogata
  6. Toru Nakamura
  7. Takuji Torimura
  8. Hideki Iwamoto
  9. Sivakumar Ramadoss
  10. Michio Sata
  11. Victor Tsutsumi
  12. Kaori Yasuda
  13. Yumi Tomiyasu
  14. Kenichi Obayashi
  15. Kosuke Tashiro
  16. Satoru Kuhara

Corresponding author

Correspondence toTakato Ueno.

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Nakamura, A., Ueno, T., Yagi, Y. et al. Human primary cultured hepatic stellate cells can be cryopreserved.Med Mol Morphol 43, 107–115 (2010). https://doi.org/10.1007/s00795-009-0484-5

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