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| 1 | Improved cryopreservation of human hepatocytes using a new xeno free cryoprotectant solution显示文摘AIM:To optimize a xeno-free cryopreservation protocol for primary human hepatocytes.METHODS:The demand for cryopreserved hepatocytes is increasing for both clinical and research purposes.Despite several hepatocyte cryopreservation protocols being available,improvements are urgently needed.We first compared controlled rate freezing to polystyrene box freezing and did not find any significant change between the groups.Using the polystyrene box freezing,we compared two xeno-free freezing solutions for freezing of primary human hepatocytes:a new medium(STEM-CELLBANKER,CB),which contains dimethylsulphoxide(DMSO) and anhydrous dextrose,both permeating and non-permeating cryoprotectants,and the frequently used DMSO-University of Wisconsin(DMSOUW) medium.The viability of the hepatocytes was assessed by the trypan blue exclusion method as well as a calcein-esterase based live-dead assay before and after cryopreservation.The function of the hepatocytes was evaluated before and after cryopreservation by assessing enzymatic activity of 6 major cytochrome P450 isoforms(CYPs):CYP1A2,CYP2C9,CYP2C19,CYP2D6,CYP3A4 and CYP3A7.RESULTS:The new cryoprotectant combination preserved hepatocyte viability significantly better than the standard DMSO-UW protocol(P < 0.01).There was no significant difference in viability estimation between both the trypan blue(TB) and the Live-Dead Assay methods.There was a correlation between viability of fresh hepatocytes and the difference in cell viability between CB and DMSO protocols(r 2 = 0.69) using the TB method.However,due to high within-group variability in the activities of the major CYPs,any statistical between-group differences were precluded.Cryopreservation of human hepatocytes using the cryoprotectant combination was a simple and xeno-free procedure yielding better hepatocyte viability.Thus,it may be a better alternative to the standard DMSO-UW protocol.Estimating CYP activities did not seem to be a relevant way to compare hepatocyte function between different groups due to high normal variability between different liver samples.CONCLUSION:The cryoprotectant combination may be a better alternative to the standard DMSO-UW protocol in primary human hepatocyte cryopreservation. | Mohammed Saliem Frida Holm Rosita Bergstrm Tengzelius Carl Jorns Lisa-Mari Nilsson Bo-Gran Ericzon Ewa Ellis Outi Hovatta | 2012 | World Journal of Hepatology2012，4，5： | 2 |
| 2 | The effects of insulin,and insulin-like growth factors Ⅰ and Ⅱ on human ovarian follicles in long-term culture显示文摘 | Louhio H Hovatta O Sj(o)berg J | 2000 | Mol Hum Reprod2000，6，8： | 1 |
| 3 | Birth of a healthy infant after invitro oocyte maturation and ICSI in a woman with diminished ovaeian response:case report显示文摘 | Friden B Hreinsson J Hovatta O | | 0，，09： | 1 |
| 4 | Effects of follicle-stim-ulating hormone and serum substitution on the in-vitro growth ofhuman ovarian follicles 显示文摘 | Wright C S Hovatta 0 Margara R | 1999 | Hum Reprod1999，14，6： | 1 |
| 5 | Luteal phase progesterone increases live birth rate after frozen embryo transfer显示文摘 | Bjuresten K Landgren BM Hovatta O | 2011 | Fertil Steril2011，95，2： | 1 |
| 6 | A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells显示文摘 | HOVATTA O MIKKOLA M GERTOW K | 2003 | Hum Report2003，18，： | 1 |
| 7 | A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells显示文摘 | Hovatta O Mikkola M Gertow K | 2003 | Hum Reprod2003，18，7： | 1 |
| 8 | A culture system using human foreskin fibroblast as feeder cells allows production of human embryonic stem cells显示文摘 | Mikkola M Gertow | 2003 | Hum Reprod2003，18，7： | 1 |
| 9 | Cultures of human embryonic stem cells: serum replacement medium or serum-containing media and the effect of basic fibroblast growth factor显示文摘 | Heidi Koivisto Marjukka Hyv?rinen Anne-Marie Str?mberg Jose Inzunza Eija Matilainen Milla Mikkola Outi Hovatta Heli Teerijoki | 2004 | Reproductive BioMedicine Online2004，，3： | 1 |
| 10 | Cryopreservation of testicular tissue显示文摘 | Hovatta | 2000 | Molecular and Cellular Endocrinology2000，169，： | 1 |
| 11 | Chromosome 1 loci in Finnish schizophrenia families显示文摘 | Ekelund J Hovatta I Parker A | 2001 | Hum Mol Genet2001，10，15： | 1 |
| 12 | Pregnancy resulting from intracytoplasmic injection of spermatozoa from a frozenthawed testicular biopsy specimen显示文摘 | Hovatta O Foudila T Siegberg R | 1996 | Hum Reprod1996，11，： | 1 |
| 13 | A culture system using human foreskin fibroblasts as feeder ceils allows production of human embryonic stem cells 显示文摘 | Hovatta O Mikkola M Gertow K | 2003 | Hum Reprod2003，18，7： | 1 |
| 14 | Clinical as-pects of fertility preservation in female patients 显示文摘 | Rodriguez - Macias Wallberg K Keros V Hovatta O | 2009 | Pediatr BloodCancer2009，53，2： | 1 |
| 15 | A culture system using human foreskin fibroblasts as feeder cells allows pro- duction of human embryonic stem cells 显示文摘 | Hovatta O Mikkolz M Gertow K | 2003 | Hum Reprod2003，18，7： | 1 |
| 16 | Germ cell apoptosis after treatment of eryptorehidism with human ehorionie gonadotropin is associated with Impaired reproductive function in the adult 显示文摘 | Leo Dunkel Seppo Taskinen Outi Hovatta | 1997 | J Clin Invest1997，100，9： | 1 |
| 17 | Methods for cryopreservation of human ovarian tissue显示文摘 | Hovatta O | | 0，，06： | 1 |
| 18 | Effects of follicle-stimulating hormone and serum substitution on the in vitro growth of human ovarian follicles 显示文摘 | WRIGHT C S HOVATTA O MARGARA R | 1999 | Hum Reprod1999，14，6： | 1 |
| 19 | A culture system using human foreskin fibroblasts as feeder cells allows production of human embryonic stem cells显示文摘 | Hovatta O Mikkola M Gertow K | 2003 | Hum Reprod2003，18，4： | 1 |
| 20 | Clinical aspects of fertility preservation in female patients 显示文摘 | Wallberg KA Keros V Hovatta O | 2009 | Pediatr Blood Cancer2009，53，： | 1 |
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