An In Vitro Mechanism Study on the Proliferation and Pluripotency of Human Embryonic Stems Cells in Response to Magnesium Degradation
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{"title"=>"An In Vitro Mechanism Study on the Proliferation and Pluripotency of Human Embryonic Stems Cells in Response to Magnesium Degradation", "type"=>"journal", "authors"=>[{"first_name"=>"Thanh Yen", "last_name"=>"Nguyen", "scopus_author_id"=>"55279555200"}, {"first_name"=>"Chee Gee", "last_name"=>"Liew", "scopus_author_id"=>"9534099800"}, {"first_name"=>"Huinan", "last_name"=>"Liu", "scopus_author_id"=>"8569854200"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"370046519", "doi"=>"10.1371/journal.pone.0076547", "sgr"=>"84885739421", "scopus"=>"2-s2.0-84885739421", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"24146887"}, "id"=>"9439eaf4-4fc4-3bba-85ca-d5096b659651", "abstract"=>"Magnesium (Mg) is a promising biodegradable metallic material for applications in cellular/tissue engineering and biomedical implants/devices. To advance clinical translation of Mg-based biomaterials, we investigated the effects and mechanisms of Mg degradation on the proliferation and pluripotency of human embryonic stem cells (hESCs). We used hESCs as the in vitro model system to study cellular responses to Mg degradation because they are sensitive to toxicants and capable of differentiating into any cell types of interest for regenerative medicine. In a previous study when hESCs were cultured in vitro with either polished metallic Mg (99.9% purity) or pre-degraded Mg, cell death was observed within the first 30 hours of culture. Excess Mg ions and hydroxide ions induced by Mg degradation may have been the causes for the observed cell death; hence, their respective effects on hESCs were investigated for the first time to reveal the potential mechanisms. For this purpose, the mTeSR®1 hESC culture media was either modified to an alkaline pH of 8.1 or supplemented with 0.4-40 mM of Mg ions. We showed that the initial increase of media pH to 8.1 had no adverse effect on hESC proliferation. At all tested Mg ion dosages, the hESCs grew to confluency and retained pluripotency as indicated by the expression of OCT4, SSEA3, and SOX2. When the supplemental Mg ion dosages increased to greater than 10 mM, however, hESC colony morphology changed and cell counts decreased. These results suggest that Mg-based implants or scaffolds are promising in combination with hESCs for regenerative medicine applications, providing their degradation rate is moderate. Additionally, the hESC culture system could serve as a standard model for cytocompatibility studies of Mg in vitro, and an identified 10 mM critical dosage of Mg ions could serve as a design guideline for safe degradation of Mg-based implants/scaffolds.", "link"=>"http://www.mendeley.com/research/vitro-mechanism-study-proliferation-pluripotency-human-embryonic-stems-cells-response-magnesium-degr", "reader_count"=>21, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>6, "Student > Master"=>1, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>6, "Student > Ph. D. Student"=>6, "Student > Master"=>1, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>4, "Biochemistry, Genetics and Molecular Biology"=>4, "Materials Science"=>2, "Agricultural and Biological Sciences"=>8, "Medicine and Dentistry"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>4}, "Materials Science"=>{"Materials Science"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}}, "group_count"=>4}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1246467"], "description"=>"<p>The ion concentrations of inorganic salts in the culture media in comparison with human blood plasma <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076547#pone.0076547-Ludwig1\" target=\"_blank\">[25]</a>, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076547#pone.0076547-Iskandar1\" target=\"_blank\">[28]</a>.</p>", "links"=>[], "tags"=>["ion", "concentrations", "inorganic", "salts", "plasma"], "article_id"=>826715, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.t002", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_ion_concentrations_of_inorganic_salts_in_the_culture_media_in_comparison_with_human_blood_plasma_25_28_/826715", "title"=>"The ion concentrations of inorganic salts in the culture media in comparison with human blood plasma [25], [28].", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246466"], "description"=>"<p>Genes screened by Q-PCR to determine pluripotency of hESCs.</p>", "links"=>[], "tags"=>["screened", "q-pcr", "pluripotency"], "article_id"=>826714, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.t001", "stats"=>{"downloads"=>3, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Genes_screened_by_Q_PCR_to_determine_pluripotency_of_hESCs_/826714", "title"=>"Genes screened by Q-PCR to determine pluripotency of hESCs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246456"], "description"=>"<p>The hESC colonies showed a tightly packed morphology in the media supplemented with 4 = 100 µm.</p>", "links"=>[], "tags"=>["optical", "images", "hescs", "cultured", "supplemented", "mg", "ion"], "article_id"=>826709, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phase_contrast_optical_images_of_hESCs_cultured_in_the_media_supplemented_with_Mg_ion_dosages_/826709", "title"=>"Phase contrast optical images of hESCs cultured in the media supplemented with Mg ion dosages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246453"], "description"=>"<p>(A) The coverage area of viable hESC colonies at 6–30 hours of culture after being normalized over the first time point at 6 hours. Values are mean ± standard error of the mean: n = 6. The hESCs cultured under the alkaline media condition proliferated and grew to confluency, indicating that the initial increase of media pH to 8.1 was not the direct cause of cell death observed in the cultures with the Mg samples. (B) The average pH of post-culture media collected at every 24 hours for 72 hours (*<i>p</i><0.05 compared to the control). Values are mean ± standard deviation: n = 3.</p>", "links"=>[], "tags"=>["viable", "hesc", "colonies", "ph", "post-culture", "alkaline"], "article_id"=>826708, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g004", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Coverage_area_of_viable_hESC_colonies_and_pH_of_post_culture_media_under_the_alkaline_or_normal_media_conditions_/826708", "title"=>"Coverage area of viable hESC colonies and pH of post-culture media under the alkaline or normal media conditions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246445"], "description"=>"<p>(A) SEM image and (B) EDS analysis of the D-Mg sample, showing the presence of a degradation layer on the surface. Accelerating voltage: 15 kV. Original Magnification: 2000x. Scale bar = 20 µm.</p>", "links"=>[], "tags"=>["morphology"], "article_id"=>826705, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Surface_morphology_and_composition_of_D_Mg_/826705", "title"=>"Surface morphology and composition of D-Mg.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246465"], "description"=>"<p>H9-OCT4 hESCs were cultured for 72 hours and analyzed for expression of the corresponding markers. No statistically significant difference was detected among any of the groups. Values are mean ± standard deviation; n = 9.</p>", "links"=>[], "tags"=>["pcr", "pluripotency", "markers", "hescs", "cultured", "supplemented", "mg", "ion"], "article_id"=>826713, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g009", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantitative_PCR_analysis_of_pluripotency_markers_for_hESCs_after_being_cultured_in_the_media_supplemented_with_Mg_ion_dosages_/826713", "title"=>"Quantitative PCR analysis of pluripotency markers for hESCs after being cultured in the media supplemented with Mg ion dosages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246462"], "description"=>"<p>(A) Average pH and (B) Mg ion concentrations in the post-culture media when the supplemental Mg ion dosages ranged from 0.4–40 mM (*<i>p</i><0.05 compared to the control at each time point respectively). Values are mean ± standard deviation; n = 3. The cell counts in the hESC cultures supplemented with 30 and 40 mM Mg ions were statistically lower than the cultures with lower Mg ion dosages and the control, and, therefore, less acidic metabolites formed and the pH values were statistically higher. Additionally, statistically greater Mg ion concentrations were observed in the post-culture media at the critical Mg ion concentration of 10 mM and above, as compared with the control, 0.4 mM and 4 mM media conditions at each respective time point.</p>", "links"=>[], "tags"=>["ph", "mg", "ion", "concentrations", "post-culture", "24", "hours", "hescs", "cultured", "supplemented"], "article_id"=>826711, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g007", "stats"=>{"downloads"=>4, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Media_pH_and_Mg_ion_concentrations_in_the_post_culture_media_collected_at_every_24_hours_after_hESCs_were_cultured_in_the_media_supplemented_with_Mg_ion_dosages_/826711", "title"=>"Media pH and Mg ion concentrations in the post-culture media collected at every 24 hours after hESCs were cultured in the media supplemented with Mg ion dosages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246464"], "description"=>"<p>All H9-OCT4 hESCs cultured in the media supplemented with different dosages of Mg ions expressed markers for OCT4 and SSEA3. The images for 0.4 mM media condition were not shown as they were very similar to the control. Nuclear localization of OCT4 is shown in green (left column) and SSEA3 expression on the cell surface is shown in red (middle column). The right column shows merged images of the OCT4, SSEA3, and DAPI (blue, cell nucleus). Scale bars = 100 µm.</p>", "links"=>[], "tags"=>["hesc", "oct4", "ssea3", "cultured", "supplemented", "mg", "ion"], "article_id"=>826712, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g008", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immunocytochemistry_analysis_on_the_hESC_expression_of_OCT4_and_SSEA3_after_being_cultured_in_the_media_supplemented_with_Mg_ion_dosages_/826712", "title"=>"Immunocytochemistry analysis on the hESC expression of OCT4 and SSEA3 after being cultured in the media supplemented with Mg ion dosages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246458"], "description"=>"<p>(A) The coverage area of viable hESC colonies after being normalized over the first time point at 0 hour in the cultures with respective supplemental Mg ion dosages (***<i>p</i><0.05 when comparing L-group with the H-group). (B) The average numbers of hESCs per 0.5 mm<sup>2</sup> after being cultured for 72 hours and immunostained with DAPI and pluripotency marker-SOX2 (*<i>p</i><0.05 compared to the control and <i>#p</i><0.05 compared to 40 mM). Although the normalized coverage area of viable hESC colonies was greater at supplemental Mg ion dosages of 10 mM and greater, the cell counts per unit area were actually lower. This confirmed that the supplemental Mg ion dosages of 10 mM and greater caused cell dispersion and loss of tightly packed morphology. Values are mean ± standard error of the mean: n = 6.</p>", "links"=>[], "tags"=>["viable", "hesc", "colonies", "numbers", "hescs", "cultured", "supplemented", "mg", "ion"], "article_id"=>826710, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g006", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Coverage_area_of_viable_hESC_colonies_and_the_numbers_of_hESCs_when_cultured_in_the_media_supplemented_with_Mg_ion_dosages_/826710", "title"=>"Coverage area of viable hESC colonies and the numbers of hESCs when cultured in the media supplemented with Mg ion dosages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246451"], "description"=>"<p>(A) The average pH and (B) the average magnesium ion concentrations (**<i>p</i><0.05 compared to M-Mg). Values are mean ± standard deviation: n = 3. The average pH and Mg ion concentrations for the culture with both Mg samples significantly increased as compared with the control without Mg samples. The statistically significant lower pH of the post-culture media with D-Mg was detected in comparison with M-Mg. Furthermore, after 24 hours, a general trend of gradual decrease in average pH and magnesium ion concentrations over time was observed for both Mg samples. (**<i>p</i><0.05 compared to M-Mg). Values are mean ± standard deviation: n = 3.</p>", "links"=>[], "tags"=>["ph", "mg", "ion", "concentrations", "24", "hours", "hescs", "cultured", "m-mg", "d-mg", "72"], "article_id"=>826707, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Media_pH_and_Mg_ion_concentrations_in_the_media_collected_at_every_24_hours_after_hESCs_were_cultured_with_M_Mg_and_D_Mg_for_72_hours_/826707", "title"=>"Media pH and Mg ion concentrations in the media collected at every 24 hours after hESCs were cultured with M-Mg and D-Mg for 72 hours.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1246449"], "description"=>"<p>As compared with the control, the H9-OCT4 hESCs exposed to the M-Mg and D-Mg showed increased dispersion at 12 and 18 hours of culture, and thus greater coverage area of cell colonies. Scale bar = 100 µm.</p>", "links"=>[], "tags"=>["optical", "images", "hesc", "colonies", "cultured", "m-mg"], "article_id"=>826706, "categories"=>["Biological Sciences"], "users"=>["Thanh Yen Nguyen", "Chee Gee Liew", "Huinan Liu"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0076547.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phase_contrast_optical_images_of_hESC_colonies_cultured_with_M_Mg_and_D_Mg_/826706", "title"=>"Phase contrast optical images of hESC colonies cultured with M-Mg and D-Mg.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-17 04:06:07"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Engineering and technology", "average_usage"=>[254, 440, 558, 675, 785, 883, 972, 1061, 1154, 1248, 1336, 1414, 1489]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[264, 460, 584, 692, 794, 887, 978, 1067, 1154, 1241, 1328, 1408, 1474]}, {"subject_area"=>"/Physical sciences", "average_usage"=>[254, 431, 547, 651, 748, 842, 932, 1017, 1098, 1178, 1259, 1336, 1404]}, {"subject_area"=>"/Physical sciences/Chemistry", "average_usage"=>[247, 429, 544, 647, 747, 842, 929, 1012, 1099, 1179, 1263, 1339, 1409]}]}
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