細胞層級的高通量篩檢系統，有助於早期藥物開發階段，快速評估上千種化合物特性。然而，這樣的篩選通常依賴於高度增殖或癌變的細胞株，在正常人類細胞中不一定能準確顯示出受影響的化合物。初代細胞 (primary cell) 的使用相當常見，但是它們有使用的限制且批次間變異性大。
人類 iPS 細胞是一個有可靠的正常人類細胞的來源，因為 iPS 細胞具有自我更新潛能和多能性分化能力，幾乎可分化成任何類型的體細胞。ReproCELL 建立了專有技術，以提供人類 iPS 細胞來源的肝細胞 (ReproHepato)，無數量限制地生產高品質、完全分化的肝細胞，使得高通量篩選僅有微小的批次變化。
ReproCELL 肝細胞優點
- 全球第一個商品化，從 iPS 細胞衍化出具有 CYP 活性的人類肝細胞
- 對照組為初代細胞 (primary cells) 時，有穩定的 CYP 活性和誘導力 (3A4/1A2/2C9/2C19) 測試結果，培養中細胞壽命長
- 細胞為已分化細胞，使用套組中的 Maitenance Medium 培養六天後，該細胞則可進行 CYP assay 試驗
- 細胞需儲存於 – 80℃，液態氮可能會降低其活性
- 帶有高活性 CYP22D6P 的產品已被研發生產
- 另提供 CYP assay 合約式檢測服務
人類 iPS 細胞衍生之肝細胞：高純度且具有 CYP 活性
ReproHepato 細胞帶有典型的人類肝細胞外型和特徵，表現肝細胞組織專一性蛋白，如 CYP3A4、HNF4a、E-cadherin，或高比例的白蛋白 (albumin) 。
明視野影像
PAS 染色：顯示肝醣囤積
免疫染色 :
G / E-cadherin
R / HNF4a
流式細胞儀分析: CYP3A4 (+) 細胞
流式細胞儀分析：帶有白蛋白細胞
穩定的 CYP 和誘導力測試結果
如同原代肝細胞，CYP3A4 的活性是通過典型的誘導劑和抑製劑以劑量效應的方式調控，其他 CYP 細胞色素（1A2/2C9/2C19）的活性和誘導力，被觀察到具有和原代肝細胞相同水平的反應。現在正開發中即將推出的 ReproHepato 版本中，還觀察到 CYP2D6 活性。
更穩定的實驗結果：批次間差異小
相較於初代肝細胞 (primary hepatocytes)，ReproHepato 肝細胞在 CYP 活性和誘導力測試中，批次間差異較低，品質穩定。
利用冷光 (luminescence) 進行 基態和 rifampicin 誘導 CYP3A4 活性測試
不同批次肝細胞 CYP3A4 活性變化比較
ReproCELL 肝細胞
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| 產品貨號 | 產品名稱 | 產品數量 | 產品分類 |
|---|---|---|---|
| RCDH001N | ReproHepato Human iPS Cell-derived Hepatocytes | 8.25×106 cells/vial (frozen cells, for plating one whole 96-well plate) | Kit |
| RCDH101 | ReproHepato Culture Medium | Basal medium: 80 mL Supplement: 400 µL | Reagents |
| RCDH301 | ReproHepato Assay Medium | Reagents | |
| RCESDH401 | CYP Assay Service | Reagents |
- Inamura, Mitsuru and Yokoyama, Chikafumi. “Human iPS cell-derived hepatocytes for drug screening.” Regenerative Medicine (2013)
- Scott, Clay W., Matthew F. Peters, and Yvonne P. Dragan. “Human induced pluripotent stem cells and their use in drug discovery for toxicity testing.” Toxicology letters 219.1 (2013): 49-58.
Additional Publications
- Murayama Nl Yamazaki H. Cytochrome P450-dependent drug oxidation activities in commercially available hepatocytes derived from human induced pluripotent stem cells cultured for 3 weeks. J Tox Sci 43:241-245 (2018).
- Kanamori T; Iwata YT; Segawa H; Yamamuro T; Kuwayama K; Tsujikawa K; Inoue H. Metabolism of fentanyl and acetylfentanyl in human-induced pluripotent stem cell-derived hapatocytes._Biol Chem Bull_41:106 (2018)
Presentations
- Society for Biomolecular Sciences(SBS), 14th AnnualConference and Exhibition 2008 (St. Louis), poster session, April 2008.
- IBC AsiaÕs 4th Annual Stem Cells Asia Congress (Singapore), oral session, June 2008.
- ELRIG and SBS Present: Drug Discovery (UK)poster session, September 2008.
- Advances in Stem Cell Discoveries (San Francisco) oral session, September 2008.
- Stem Cells: Drug Discovery and Therapeutics (London) oral session, February 2008.
- Society for Biomolecular Science 15th Annual Conference (France), poster session, Best Poster 2009 Award-winning, April 2009.
- Stem Cells and Regenerative Medicine Europe (UK), September 2009,Best Poster 2009 Award-winning.
- MipTec Conference 2009, October 13-15, Basel Invited Lecture.
- The 74th Annual Scientific Meeting of the Japanese Circulation Society (Kyoto), March 2010.
- The 130th The Pharmaceutical Society of Japan (Okayama),March 2010.
- The Society of Toxicology (SOT) 51st Annual Meeting (CA, USA), poster session, March 2012.
- Safety Pharmacology Society (SPS), 13th annual meeting 2013 (Rotterdam, Netherlands), poster session, September 2013.
- Automated Patch-Clamp systems and Ion Channel Expressing Cells 2013 (Tokyo), Verbal presentation 2013 Nov.
- The 7th Takeda Science Foundation Symposium (Osaka), Poster presentation 2014 Jan.
- Inamura, Mitsuru and Yokoyama, Chikafumi. “Human iPS cell-derived hepatocytes for drug screening.” Regenerative Medicine (2013)
- Scott, Clay W., Matthew F. Peters, and Yvonne P. Dragan. “Human induced pluripotent stem cells and their use in drug discovery for toxicity testing.” Toxicology letters 219.1 (2013): 49-58.
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