High concentration of synthetic serum, stepwise equilibration and slow cooling as an efficient technique for large-scale cryopreservation of human embryonic stem cells

2010 ◽  
Vol 93 (3) ◽  
pp. 976-985 ◽  
Author(s):  
Ji Yeon Lee ◽  
Jeoung Eun Lee ◽  
Dong Ku Kim ◽  
Tae Ki Yoon ◽  
Hyung Min Chung ◽  
...  
Cryobiology ◽  
2006 ◽  
Vol 53 (2) ◽  
pp. 194-205 ◽  
Author(s):  
Igor I. Katkov ◽  
Min S. Kim ◽  
Ruchi Bajpai ◽  
Yoav S. Altman ◽  
Marc Mercola ◽  
...  

2012 ◽  
Vol 7 (2) ◽  
pp. 40-43
Author(s):  
T Hasan

Introduction: Human-embryonic stem cells (hESC) are derived from very early stages of the human embryo. These cells have immense plasticity and can be conditioned to develop into any type of cell of the human body. Despite all their promising utility, hESC researches have recently been the subject of fervent debate. Objective: This paper explores the implications of hESC therapy from a bio-ethical perspective. Method: Published literature with strict inclusion and exclusion criteria was extensively reviewed through use of general and meta search engines to elucidate the applications and implications of hESC. Discussion: Studies indicate that the potential of hESC in reconstructive and regenerative medicine is undisputable but complex social and moral issues are hopelessly intertwined beneath the pleasant facade. hESC offer endless possibilities in understanding bio-molecular disease patterns, supplying readymade healthy organs, interpreting aging and organogenesis at the cellular level. The use of hESC is well established in leukemia and scientists anticipate diverse applications in a wide range of congenital and acquired medical conditions. However, many dilemmas arise in context of their biomedical usage because of the destruction of donor human embryos in producing stem cells, adverse transplant reactions, teratogenecity, phenotypic / genotypic abnormalities, nonstandardized research laws, logistic issues and the possibility of eternal life and humanoid chimeras. Conclusion: The wisdom to choose between ' mindful utilization' and 'senseless exploitation' lies with us. The large scale commercialization of human life or the killing of viable embryos cannot be justified by any means. A neutral approach with increased involvement of uncontroversial progenitors should be adopted. DOI: http://dx.doi.org/10.3329/jafmc.v7i2.10396 JAFMC 2011; 7(2): 40-43


2007 ◽  
Vol 27 (4-5) ◽  
pp. 257-264 ◽  
Author(s):  
Boon Chin Heng ◽  
Marie Veronique Clement ◽  
Tong Cao

Previous study demonstrated that the low survival of human embryonic stem cells (hESC) under conventional slow-cooling cryopreservation protocols is predominantly due to apoptosis rather than cellular necrosis. Hence, this study investigated whether a synthetic broad-spectrum irreversible inhibitor of caspase enzymes, Z-VAD-FMK can be used to enhance the post-thaw survival rate of hESC. About 100 mM Z-VAD-FMK was supplemented into either the freezing solution, the post-thaw culture media or both. Intact and adherent hESC colonies were cryopreserved so as to enable subsequent quantitation of the post-thaw cell survival rate through the MTT assay, which can only be performed with adherent cells. Exposure to 100 mM Z-VAD-FMK in the freezing solution alone did not significantly enhance the post-thaw survival rate (10.2% vs. 9.9%, p > 0.05). However, when 100 mM Z-VAD-FMK was added to the post-thaw culture media, there was a significant enhancement in the survival rate from 9.9% to 14.4% (p < 0.05), which was further increased to 18.7% when Z-VAD-FMK was also added to the freezing solution as well (p < 0.01). Spontaneous differentiation of hESC after cryopreservation was assessed by morphological observations under bright-field microscopy, and by immunocytochemical staining for the pluripotency markers SSEA-3 and TRA-1-81. The results demonstrated that exposure to Z-VAD-FMK did not significantly enhance the spontaneous differentiation of hESC within post-thaw culture.


2010 ◽  
Vol 88 (16) ◽  
pp. 3467-3478 ◽  
Author(s):  
Asuka Morizane ◽  
Vladimer Darsalia ◽  
M. Oktar Guloglu ◽  
Tord Hjalt ◽  
Manolo Carta ◽  
...  

BMC Genomics ◽  
2007 ◽  
Vol 8 (1) ◽  
pp. 478 ◽  
Author(s):  
Thomas C Schulz ◽  
Anna Swistowska ◽  
Ying Liu ◽  
Andrzej Swistowski ◽  
Gail Palmarini ◽  
...  

Blood ◽  
2008 ◽  
Vol 112 (12) ◽  
pp. 4475-4484 ◽  
Author(s):  
Shi-Jiang Lu ◽  
Qiang Feng ◽  
Jennifer S. Park ◽  
Loyda Vida ◽  
Bao-Shiang Lee ◽  
...  

Abstract Human erythropoiesis is a complex multistep process that involves the differentiation of early erythroid progenitors to mature erythrocytes. Here we show that it is feasible to differentiate and mature human embryonic stem cells (hESCs) into functional oxygen-carrying erythrocytes on a large scale (1010-1011 cells/6-well plate hESCs). We also show for the first time that the oxygen equilibrium curves of the hESC-derived cells are comparable with normal red blood cells and respond to changes in pH and 2,3-diphosphoglyerate. Although these cells mainly expressed fetal and embryonic globins, they also possessed the capacity to express the adult β-globin chain on further maturation in vitro. Polymerase chain reaction and globin chain specific immunofluorescent analysis showed that the cells increased expression of β-globin (from 0% to > 16%) after in vitro culture. Importantly, the cells underwent multiple maturation events, including a progressive decrease in size, increase in glycophorin A expression, and chromatin and nuclear condensation. This process resulted in extrusion of the pycnotic nuclei in up to more than 60% of the cells generating red blood cells with a diameter of approximately 6 to 8 μm. The results show that it is feasible to differentiate and mature hESCs into functional oxygen-carrying erythrocytes on a large scale.


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