Islet-Like Cell Aggregates Generated from Human Adipose Tissue Derived Stem Cells Ameliorate Experimental Diabetes in Mice
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{"title"=>"Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice", "type"=>"journal", "authors"=>[{"first_name"=>"Vikash", "last_name"=>"Chandra", "scopus_author_id"=>"14818872500"}, {"first_name"=>"Swetha", "last_name"=>"G", "scopus_author_id"=>"35336315700"}, {"first_name"=>"Sudhakar", "last_name"=>"Muthyala", "scopus_author_id"=>"36619110100"}, {"first_name"=>"Amit K.", "last_name"=>"Jaiswal", "scopus_author_id"=>"55444791600"}, {"first_name"=>"Jayesh R.", "last_name"=>"Bellare", "scopus_author_id"=>"7003688762"}, {"first_name"=>"Prabha D.", "last_name"=>"Nair", "scopus_author_id"=>"7201717296"}, {"first_name"=>"Ramesh R.", "last_name"=>"Bhonde", "scopus_author_id"=>"7005467058"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"361908276", "sgr"=>"79958139078", "issn"=>"19326203", "pmid"=>"21687731", "scopus"=>"2-s2.0-79958139078", "doi"=>"10.1371/journal.pone.0020615", "isbn"=>"1932-6203"}, "id"=>"1ff60d9d-fdb1-31be-87ae-0db990f4c4d2", "abstract"=>"BACKGROUND: Type 1 Diabetes Mellitus is caused by auto immune destruction of insulin producing beta cells in the pancreas. Currently available treatments include transplantation of isolated islets from donor pancreas to the patient. However, this method is limited by inadequate means of immuno-suppression to prevent islet rejection and importantly, limited supply of islets for transplantation. Autologous adult stem cells are now considered for cell replacement therapy in diabetes as it has the potential to generate neo-islets which are genetically part of the treated individual. Adopting methods of islet encapsulation in immuno-isolatory devices would eliminate the need for immuno-suppressants. METHODOLOGY/PRINCIPAL FINDINGS: In the present study we explore the potential of human adipose tissue derived adult stem cells (h-ASCs) to differentiate into functional islet like cell aggregates (ICAs). Our stage specific differentiation protocol permit the conversion of mesodermic h-ASCs to definitive endoderm (Hnf3beta, TCF2 and Sox17) and to PDX1, Ngn3, NeuroD, Pax4 positive pancreatic endoderm which further matures in vitro to secrete insulin. These ICAs are shown to produce human C-peptide in a glucose dependent manner exhibiting in-vitro functionality. Transplantation of mature ICAs, packed in immuno-isolatory biocompatible capsules to STZ induced diabetic mice restored near normoglycemia within 3-4 weeks. The detection of human C-peptide, 1155+/-165 pM in blood serum of experimental mice demonstrate the efficacy of our differentiation approach. CONCLUSIONS: h-ASC is an ideal population of personal stem cells for cell replacement therapy, given that they are abundant, easily available and autologous in origin. Our findings present evidence that h-ASCs could be induced to differentiate into physiologically competent functional islet like cell aggregates, which may provide as a source of alternative islets for cell replacement therapy in type 1 diabetes.", "link"=>"http://www.mendeley.com/research/isletlike-cell-aggregates-generated-human-adipose-tissue-derived-stem-cells-ameliorate-experimental", "reader_count"=>9, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>3, "Student > Master"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>3, "Student > Master"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>1, "Medicine and Dentistry"=>2, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/768337"], "description"=>"<p>qRT PCR based transcript abundance of pancreatic hormones (Insulin, Glucagon, Somatostatin, Pan-pol and Ghrelin) in undifferentiated and day 14 differentiated h-ASCs. Transcript abundance of these genes was also compared to that of human fetal pancreas 12 week (H-Pan). Relative levels of gene expression were normalized to the GAPDH mRNA level and presented here as fold increase of mean±S.E.M over detectable (Ct of 39) (A). (White bar: Undifferentiated h-ASCs, Grey bar: Differentiated ICAs on day 14 and Green bar: Human fetal pancreas- 12 week). Scanning electron microscopy (SEM) of day 14 ICAs show typical surface topography similar to that of pancreatic islets. Scale bar = 200 µm (B) and 100 µm (C). Abbreviations: qRT PCR, quantitative reverse transcription polymerase chain reaction; cDNA, complementary DNA; pan pol, pancreatic polypeptide; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase.</p>", "links"=>[], "tags"=>["pancreatic", "endocrine", "hormones", "transcript", "topography", "14", "icas", "scanning", "electron"], "article_id"=>438688, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_pancreatic_endocrine_hormones_transcript_during_the_course_of_differentiation_surface_topography_of_day_14_ICAS_with_scanning_electron_microscopy_/438688", "title"=>"Expression of pancreatic endocrine hormones transcript during the course of differentiation, surface topography of day 14 ICAS with scanning electron microscopy.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:24:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/385846", "https://ndownloader.figshare.com/files/385859", "https://ndownloader.figshare.com/files/385872", "https://ndownloader.figshare.com/files/385886", "https://ndownloader.figshare.com/files/385893"], "description"=>"<div><h3>Background</h3><p>Type 1 Diabetes Mellitus is caused by auto immune destruction of insulin producing beta cells in the pancreas. Currently available treatments include transplantation of isolated islets from donor pancreas to the patient. However, this method is limited by inadequate means of immuno-suppression to prevent islet rejection and importantly, limited supply of islets for transplantation. Autologous adult stem cells are now considered for cell replacement therapy in diabetes as it has the potential to generate neo-islets which are genetically part of the treated individual. Adopting methods of islet encapsulation in immuno-isolatory devices would eliminate the need for immuno-suppressants.</p> <h3>Methodology/Principal Findings</h3><p>In the present study we explore the potential of human adipose tissue derived adult stem cells (h-ASCs) to differentiate into functional islet like cell aggregates (ICAs). Our stage specific differentiation protocol permit the conversion of mesodermic h-ASCs to definitive endoderm (Hnf3β, TCF2 and Sox17) and to PDX1, Ngn3, NeuroD, Pax4 positive pancreatic endoderm which further matures in vitro to secrete insulin. These ICAs are shown to produce human C-peptide in a glucose dependent manner exhibiting in-vitro functionality. Transplantation of mature ICAs, packed in immuno-isolatory biocompatible capsules to STZ induced diabetic mice restored near normoglycemia within 3–4 weeks. The detection of human C-peptide, 1155±165 pM in blood serum of experimental mice demonstrate the efficacy of our differentiation approach.</p> <h3>Conclusions</h3><p>h-ASC is an ideal population of personal stem cells for cell replacement therapy, given that they are abundant, easily available and autologous in origin. Our findings present evidence that h-ASCs could be induced to differentiate into physiologically competent functional islet like cell aggregates, which may provide as a source of alternative islets for cell replacement therapy in type 1 diabetes.</p> </div>", "links"=>[], "tags"=>["islet-like", "aggregates", "generated", "adipose", "derived", "cells", "ameliorate", "mice"], "article_id"=>136167, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0020615.s001", "https://dx.doi.org/10.1371/journal.pone.0020615.s002", "https://dx.doi.org/10.1371/journal.pone.0020615.s003", "https://dx.doi.org/10.1371/journal.pone.0020615.s004", "https://dx.doi.org/10.1371/journal.pone.0020615.s005"], "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Islet_Like_Cell_Aggregates_Generated_from_Human_Adipose_Tissue_Derived_Stem_Cells_Ameliorate_Experimental_Diabetes_in_Mice/136167", "title"=>"Islet-Like Cell Aggregates Generated from Human Adipose Tissue Derived Stem Cells Ameliorate Experimental Diabetes in Mice", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-06-07 01:42:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/768496"], "description"=>"<p>Confocal optical sections of ICAs stained for PDX-1(a), C-peptide (b), Insulin (c), Somatostatin (d), and Glut 2 (e). Nuclei are stained by DAPI (4′, 6-diamidoino-2-phenylindole). (Scale bar = 20 µm).</p>", "links"=>[], "tags"=>["immunocytochemistry", "day14", "icas", "shows", "pancreatic", "endoderm"], "article_id"=>438856, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g007", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Confocal_immunocytochemistry_of_day14_ICAs_shows_the_expression_of_pancreatic_endoderm_markers_/438856", "title"=>"Confocal immunocytochemistry of day14 ICAs shows the expression of pancreatic endoderm markers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:27:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/767930"], "description"=>"<p>h-ASCs in monolayer when exposed to SFM A undergo cell clustering and aggregation. Subsequent exposure and incubation of h-ASCs in SFM B on day 3 and SFM C on day 5 results in the formation of mature Islet like cell aggregates (ICAs). By day14, the ICAs become fully differentiated and mature and stain positive for DTZ. Abbreviations: SFM, Serum free media; ITS, Insulin-transferrin-selenium; BSA, Bovine serum albumin, GLP- Glucagon Like Peptide, DTZ, dithizone.</p>", "links"=>[], "tags"=>["h-ascs", "derived", "14", "icas", "stained", "islet", "dtz"], "article_id"=>438287, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Differentiation_strategy_for_h_ASCs_derived_ICAs_day_14_ICAs_stained_positive_with_islet_specific_DTZ_staining_/438287", "title"=>"Differentiation strategy for h-ASCs derived ICAs, day 14 ICAs stained positive with islet specific DTZ staining.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:18:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/767754"], "description"=>"<p>Representative immunostaining of cytoskeletal protein vimentin (A), alpha smooth muscle actin (B), nestin (C), fibronectin (D) and cell proliferation marker Ki-67 (E) with immunofluoresence staining. Negative control staining with fluorescence conjugated secondary antibodies shown (F). Cell nuclei were stained with DAPI. Scale bar, 20 µm. h-ASCs exhibited competence to differentiate into adipogenic, osteogenic and chondrogenic lineages in-vitro upon specific induction as confirmed by Oil red O (G), Saffranin-O (H) and Alizarin-red (I) staining respectively.</p>", "links"=>[], "tags"=>["vitro", "characterization", "clonally", "expanded", "h-ascs", "mesenchymal"], "article_id"=>438113, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g002", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_characterization_of_clonally_expanded_h_ASCs_for_mesenchymal_markers_/438113", "title"=>"In vitro characterization of clonally expanded h-ASCs for mesenchymal markers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:15:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/768191"], "description"=>"<p>h-ASCs when differentiated to mature ICAs expressed various genes involved in pancreas development. SYBR-Green based qRT PCR was performed for the pancreatic progenitor specific transcription factors like PDX1, Ngn3, NeuroD, Pax4, Nkx2.2, Nkx6.1, Pax6, Isl1 and glucose transporter Glut-2 at day14 (Diff-d14) of differentiation. Gene transcripts of ICAs are compared with human fetal pancreas (12 week) (H-Pan). Relative levels of gene expression were normalized to the GAPDH mRNA level. Transcript abundance is shown here as fold increase of mean±S.E.M. for two experiments and represented as fold over undifferentiated h-ASCs. Abbreviations: Diff-d14: Differentiated ICAs on day 14 and H-Pan: Human fetal pancreas- 12 week).</p>", "links"=>[], "tags"=>["pancreatic", "endoderm", "genes"], "article_id"=>438557, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_profile_of_pancreatic_endoderm_genes_during_the_course_of_differentiation_/438557", "title"=>"Expression profile of pancreatic endoderm genes during the course of differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:22:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/767604"], "description"=>"<p>Flow cytometry based characterization of the clonal population of h-ASCs. Representative profile of clonally expanded h-ASCs (n = 4); cells are triple positive for CD90, CD44 and CD29 (>90%) antigens. Percentage of positivity indicated in top right corner. Three different color conjugated antibodies were used, CD90-APC, CD44-PE and CD29-FITC. Isotype controls were used for all antibody combinations.</p>", "links"=>[], "tags"=>["co-expression", "cd90-cd29-cd44", "clonally", "expanded"], "article_id"=>437966, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g001", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Uniform_co_expression_of_CD90_CD29_CD44_in_clonally_expanded_h_ASCs_/437966", "title"=>"Uniform co-expression of CD90-CD29-CD44 in clonally expanded h-ASCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:12:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/768634"], "description"=>"<p>Static stimulation assay for day14 ICAs. ICAs were sequentially treated with low glucose (5.5 mM) and high glucose (22 mM) concentration in triplicate for one hour. Cell supernatants were collected and analyzed for c-peptide release by EIA assay (<i>p</i> values≤0.05) (n = 3) (A). Total intracellular c-peptide content of day 14 ICAs (<i>p</i>≤0.05) was measure in acid-ethanol and compared equal number of undifferentiated h-ASCs (<i>p</i>≤0.05), (n = 3) (B). In-vivo characterization of h-ASCs derived ICAs. Transplantation of day-14 ICAs in STZ induced diabetic Swiss albino mice (n = 3+3) showed hypoglycemic effect within two weeks. Day-14 ICAs (1000–1200) were encapsulated in calcium-alginate and packed into biocompatible capsules (made of PU-PVP-IPN) and transplanted intraperitoneally to diabetic mice. Control group of animals included sham control of diabetic mice transplanted with empty capsule (n = 3) and diabetic mice transplanted with encapsulated undifferentiated h-ASCs (n = 3+3). Overnight fasting blood glucose levels of mice in each experimental group for three week time period is represented (C). Quantification of human C-peptide in blood serum of mice transplanted with ICAs and diabetic control mice (D). Serum samples of experimental mice were collected by retro orbital bleeding on 28<sup>th</sup> day of transplantation, for the detection of human c-peptide/mouse c-peptide in blood with ultrasensitive human c-peptide/mouse c-peptide kit were used. Abbreviations: ICAs, Islet like cell aggregates; STZ, streptozotocin; IP-GTT, intra-peritoneal glucose tolerance test; DAPI, (4′, 6-diamidoino-2-phenylindole).</p>", "links"=>[], "tags"=>["in-vivo", "characterization"], "article_id"=>439001, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g008", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_and_in_vivo_functional_characterization_of_ICAs_/439001", "title"=>"In-vitro and in-vivo functional characterization of ICAs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:30:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/768041"], "description"=>"<p>Immuno-staining of ICAs exposed to SFM A on day-2 for the expression of definitive endoderm markers Foxa2 (HNF3B) (A-a), TCF-2 (A-b) and Sox-17 (A-c) as compared to undifferentiated h-ASCs (top panel of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0020615#pone-0020615-g004\" target=\"_blank\">Figure 4</a>). Nuclei stained with DAPI. Scales bar = 20 µm. qRT-PCR data on the expression of definitive endoderm markers Foxa2, CK-19 and Sox-17 during specific stages of differentiation (day 2, day 5 and day 14) ICAs and compared with undifferentiated h-ASCs. All mRNA expression levels were normalized to the house keeping gene GAPDH expression, data presented here as fold increase of mean±S.E.M over h-ASCs undifferentiated cells (B). Abbreviations: DAPI, (4′, 6-diamidoino-2-phenylindole); CK-19, cytokeratin-19; qRT PCR, quantitative reverse transcription polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase.</p>", "links"=>[], "tags"=>["h-ascs", "derived", "icas", "endoderm"], "article_id"=>438401, "categories"=>["Chemistry", "Cell Biology", "Developmental Biology", "Immunology"], "users"=>["Vikash Chandra", "Swetha G", "Sudhakar Muthyala", "Amit K. Jaiswal", "Jayesh R. Bellare", "Prabha D. Nair", "Ramesh R. Bhonde"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020615.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_h_ASCs_derived_ICAs_for_endoderm_markers_/438401", "title"=>"Characterization of h-ASCs derived ICAs for endoderm markers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-07 02:20:01"}

PMC Usage Stats | Further Information

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