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Description	Enhanced Primary Human Hepatocytes are expanded primary cells which retains the physiologically relevant profile and phenotype of primary hepatocytes. These cells are ready-to-use for applications in research and development, such as viral infections, screening, co-cultures, transient transfection, and for metabolism studies. The late expansion cells are tested for CYP induction and inhibition. It is not recommended for further extensive passaging after thawing but can be used for long term culture which is necessary for cell-based assays. Enhanced Primary Human Hepatocytes -1 Million is a lower cell count of our expanded primary hepatocytes offered at abm. We have a higher cell count available as the Enhanced Primary Human Hepatocytes – 5 Million (Cat. T5996).
Procedure Overview
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Liver
Growth Properties	Adherent
Morphology	Polygonal
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Recommended Seeding Density	10,000 cells/cm2
Applications	For Research Use Only
Propagation	Use of Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. We strongly recommend end users to purchase the Applied Cell Extracellular Matrix (G422) to coat your cell culture vessels. Use the ready-to-use Hepatocyte Thawing Medium (TM5995-1; 50ml) and the Enhanced Primary Human Hepatocytes Media Kit (TM5995) which comes with the Hepatocyte Growth Basal Medium (TM5995-2; 500 ml), and Hepatocyte Growth Supplement Mix (TM5995-3) available from abm . Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. To make complete Hepatocyte Growth Basal Medium add the entire content of Hepatocyte Growth Supplement Mix (TM5995-3) into Hepatocyte Growth Basal Medium (TM5995-2) and mix properly. Addition of the Hepatocyte Growth Supplement Mix may change medium appearance more opaque. To thaw: 1. Pre-warm Hepatocyte Thawing Medium and fully supplemented Hepatocyte Growth Medium to room temperature. 2. Carefully remove cryovial from storage tank. 3. Thaw cells in 37°C water bath until only a small chunk of ice is left. Do not shake the vial, or take it out of the water during thawing, as this will damage the cells. 4. Spray the vial and the tube containing 50 ml of thawing medium with 70% ethanol and transfer to a Biosafety cabinet. 5. Transfer the now completely thawed cell suspension from the cryovial into 50 ml Hepatocyte Thawing Medium by gently pouring the cells into the medium. 6. Use a 1 ml pipette to transfer 1 ml of the thawing medium back to the cryovial and pour the contents back into the 50 ml tube. Repeat this process twice to completely remove the cells from the cryovial. 7. Pellet the cells by centrifuging at 90×g for 5 min at RT. Important note: Higher g-forces will significantly reduce cell recovery. 8. Aspirate the supernatant without disturbing the pellet. Leave approximately 200-400 μl medium on top of the cells. 9. Gently loosen and re-suspend the cells without adding any extra medium by agitating and rotating the tube. Do not vortex or shake the cells as this will compromise cell survival. 10. Add an appropriate volume of pre-warmed supplemented Hepatocyte Growth Medium to the pellet (~1ml per million cells thawed) and re-suspend the cells. Avoid pipetting the cells up and down. 11. Determine viable cell number by cell count. 12. Dilute hepatocytes in pre-warmed, fully supplemented Hepatocyte Growth Medium and seed at ~10,000 cells/cm2in collagen coated cell culture flasks (e.g. T175) or appropriate cell culture dishes. 13. Incubate the cells at 95% humidity, 37°C and 5% CO2.
Subculturing	To Subculture:

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合肥星肽生物科技有限公司是一家致力于生命科學和生物技術領域的高科技企業，公司地址位于科教名城———合肥市。公司主營多肽合成、多肽定制、多肽純化等多肽制備多肽技術服務。在合肥市與中科院的關懷下，2021年初合肥市高新區與中科院分別與合肥星肽生物科技有限公司簽訂了戰略投資協議。 公司位于合肥市高新區安徽中科資城創業孵化器A區，擁有2000平方米獨立研發中心以及無菌實驗室，四條完善的多肽生產線，配備了Waters LC- MS/MS、Waters UPLC、Waters 全自動純化儀、島津LCMS、全自動凍干系統、紫外分光光度計等專用精密儀器保障合肥星肽生物科技有限公司月產3000條精品多肽的生產能力，這些得以讓我們可以為客戶快速準確的提供各種高質量的多肽產品及服務。 合肥合肥星肽生物科技有限公司始終致力于為您的實驗研究提供高品質、穩定性良好的產品，“做您身邊的多肽合成專家”一直是每個合肥星肽生物科技有限公司人的目標與方向，勵志作為多肽合成行業標準的風向標。公司擁有100余人的專業團隊，其中博士研究生、碩士研究生10余人。在此基礎上公司配備了一套包含從售前到售后的專業團隊，力爭做到每個環節都“清晰明了”，對每一個問題都“吹毛求疵”，讓您的實驗變得更加簡單，正是憑借這樣的信仰和扎實的多肽合成技術優勢，合肥星肽生物科技有限公司已經成為國內外高校與眾多生物醫藥企業誠信的合作伙伴和堅強的技術后盾。 公司秉承“專業鍛造品質，服務打造市場”的經營理念，為國內廣大客戶提供專業的前沿產品、合理的價格、便捷的物流服務。 公司自成立以來，一直致力于多肽合成、多肽定制、多肽純化等多肽制備多肽技術服務的研究。幫助客戶縮短科研周期、節省試驗成本，使客戶的科學研究具有持續性。 實驗室配備一支年輕、專業、富有創造精神且經驗豐富的技術團隊。 合肥星肽生物科技有限公司目前跟國內外企業有著友好的合作往來。斯坦福大學，牛津大學，北京大學，清華大學，天津大學，浙江大學，中科大，香港理工大學，以及中科院遺傳所，過程所，化學所，生物細胞所，微生物所等，北京軍科院**醫學研究所，武漢病毒研究所等有穩定合作。

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