{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:24:13Z","timestamp":1787019853544,"version":"build-2736575974"},"reference-count":33,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Orphanet J Rare Dis"],"published-print":{"date-parts":[[2013,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Niemann Pick C (NPC) disease is a neurovisceral lysosomal storage disorder due to mutations in\n                      <jats:italic>NPC1<\/jats:italic>\n                      or\n                      <jats:italic>NPC2<\/jats:italic>\n                      genes, characterized by the accumulation of endocytosed unesterified cholesterol, gangliosides and other lipids within the lysosomes\/late endosomes. Even if the neurodegeneration is the main feature of the disease, the analysis of the molecular pathways linking the lipid accumulation and cellular damage in the brain has been challenging due to the limited availability of human neuronal models.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>The aim of this study was to develop a human neuronal model of NPC disease by inducing neuronal differentiation of multipotent adult stem cells (MASC) isolated from NPC patients.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Stem cells were isolated from 3 NPC patients and 3 controls both from skin biopsies and previously established skin fibroblast cultures. Cells were induced to differentiate along a neuronal fate adapting methods previously described by Beltrami et al, 2007. The surface immunophenotype of stem cells was analyzed by FACS. Stem cell and neuronal markers expression were evaluated by immunofluorescence. Intracellular accumulation of cholesterol and gangliosides were assessed by filipin staining and immunofluorescence, respectively. A morphometric analysis was performed using a Neurite outgrowth image program.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>After 3 passages in selective medium, MASC isolated either from skin biopsies or previously established skin fibroblast cultures displayed an antigenic pattern characteristic of mesenchymal stem cells and expressed the stem cell markers Oct-4, Nanog, Sox-2 and nestin. A massive lysosomal accumulation of cholesterol was observed only in cells isolated from NPC patients. After the induction of neural differentiation, remarkable morphologic changes were observed and cells became positive to markers of the neuronal lineage NeuN and MAP2. Differentiated cells from NPC patients displayed characteristic features of NPC disease, they showed intracellular accumulation of unesterified cholesterol and GM2 ganglioside and presented morphological differences with respect to cells derived from healthy donors.<\/jats:p>\n                    <jats:p>In conclusion, we generated a human neuronal model of NPC disease through the induction of differentiation of stem cells obtained from patient\u2019s easily accessible sources. The strategy described here may be applied to easily generate human neuronal models of other neurodegenerative diseases.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/1750-1172-8-34","type":"journal-article","created":{"date-parts":[[2013,2,21]],"date-time":"2013-02-21T19:32:21Z","timestamp":1361475141000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["A human neuronal model of Niemann Pick C disease developed from stem cells isolated from patient\u2019s skin"],"prefix":"10.1186","volume":"8","author":[{"given":"Natascha","family":"Bergamin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Dardis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Beltrami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Cesselli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Rigo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefania","family":"Zampieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rossana","family":"Domenis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno","family":"Bembi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlo Alberto","family":"Beltrami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2013,2,21]]},"reference":[{"key":"559_CR1","first-page":"611","volume-title":"The metabolic and molecular basis of inherited diseases, Volume 3","author":"M Patterson","year":"2001","unstructured":"Patterson M, Vanier MT, Suzuki K, Morris ED, Cartsea EB, Neufeld EJ, Blanchette-Mackie PG, Pentchev : Niemann Pick disease type C: a lipid trafficking disorder. The metabolic and molecular basis of inherited diseases, Volume 3. Edited by: Scriver CR, Beaudet AL, Sly WS, Valle D. 2001, New York: Mc Graw-Hill, 611-634. VII","edition":"VII"},{"key":"559_CR2","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1034\/j.1399-0004.2003.00147.x","volume":"64","author":"MT Vanier","year":"2003","unstructured":"Vanier MT, Millat G: Niemann-Pick disease type C. Clin Genet. 2003, 64: 269-281. 10.1034\/j.1399-0004.2003.00147.x.","journal-title":"Clin Genet"},{"key":"559_CR3","doi-asserted-by":"publisher","first-page":"2002","DOI":"10.1073\/pnas.90.5.2002","volume":"90","author":"ED Carstea","year":"1993","unstructured":"Carstea ED, Polymeropoulos MH, Parker CC, Detera-Wadleigh SD, O'Neill RR, Patterson MC, Goldin E, Xiao H, Straub RE, Vanier MT, Roscoe OB, Pentchev PG: Linkage of Niemann-Pick disease type C to human chromosome 18. Proc Natl Acad Sci U S A. 1993, 90: 2002-2004. 10.1073\/pnas.90.5.2002.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"559_CR4","first-page":"118","volume":"58","author":"MT Vanier","year":"1996","unstructured":"Vanier MT, Duthel S, Rodriguez-Lafrasse C, Pentchev P, Carstea ED: Genetic heterogeneity in Niemann-Pick C disease: a study using somatic cell hybridization and linkage analysis. Am J Hum Genet. 1996, 58: 118-125.","journal-title":"Am J Hum Genet"},{"key":"559_CR5","doi-asserted-by":"publisher","first-page":"24367","DOI":"10.1074\/jbc.M002184200","volume":"275","author":"JP Davies","year":"2000","unstructured":"Davies JP, Ioannou YA: Topological analysis of Niemann-Pick C1 protein reveals that the membrane orientation of the putative sterol-sensing domain is identical to those of 3-hydroxy-3-methylglutaryl-CoA reductase and sterol regulatory element binding protein cleavage-activating protein. J Biol Chem. 2000, 275: 24367-24374. 10.1074\/jbc.M002184200.","journal-title":"J Biol Chem"},{"key":"559_CR6","doi-asserted-by":"publisher","first-page":"2298","DOI":"10.1126\/science.290.5500.2298","volume":"290","author":"S Naureckiene","year":"2000","unstructured":"Naureckiene S, Sleat DE, Lackland H, Fensom A, Vanier MT, Wattiaux R, Jadot M, Lobel P: Identification of HE1 as the second gene of Niemann-Pick C disease. Science. 2000, 290: 2298-2301. 10.1126\/science.290.5500.2298.","journal-title":"Science"},{"key":"559_CR7","doi-asserted-by":"crossref","first-page":"5784","DOI":"10.1016\/S0021-9258(18)91082-3","volume":"259","author":"AD Boothe","year":"1984","unstructured":"Boothe AD, Kruth HS, Weintroub H, Stivers J, Brady RO: A genetic storage disorder in BALB\/C mice with a metabolic block in esterification of exogenous cholesterol. J Biol Chem. 1984, 259: 5784-5791.","journal-title":"J Biol Chem"},{"key":"559_CR8","doi-asserted-by":"publisher","first-page":"730","DOI":"10.1093\/hmg\/ddr505","volume":"21","author":"RA Maue","year":"2012","unstructured":"Maue RA, Burgess RW, Wang B, Wooley CM, Seburn KL, Vanier MT, Rogers MA, Chang CC, Chang TY, Harris BT, Graber DJ, Penatti CA, Porter DM, Szwergold BS, Henderson LP, Totenhagen JW, Trouard TP, Borbon IA, Erickson RP: A novel mouse model of Niemann-Pick type C disease carrying a D1005G-Npc1 mutation comparable to commonly observed human mutations. Hum Mol Genet. 2012, 21: 730-750. 10.1093\/hmg\/ddr505.","journal-title":"Hum Mol Genet"},{"key":"559_CR9","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1093\/brain\/118.1.119","volume":"118","author":"S Love","year":"1995","unstructured":"Love S, Bridges LR, Case CP: Neurofibrillary tangles in Niemann-Pick disease type C. Brain. 1995, 118: 119-129. 10.1093\/brain\/118.1.119.","journal-title":"Brain"},{"key":"559_CR10","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1016\/j.bbalip.2004.08.011","volume":"1685","author":"SU Walkley","year":"2004","unstructured":"Walkley SU, Suzuki K: Consequences of NPC1 and NPC2 loss of function in mammalian neurons. Biochim Biophys Acta. 2004, 1685: 48-62. 10.1016\/j.bbalip.2004.08.011.","journal-title":"Biochim Biophys Acta"},{"key":"559_CR11","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1042\/BST0340399","volume":"34","author":"JE Vance","year":"2006","unstructured":"Vance JE, Karten B, Hayashi H: Lipid dynamics in neurons. Biochem Soc Trans. 2006, 34: 399-403. 10.1042\/BST0340399.","journal-title":"Biochem Soc Trans"},{"key":"559_CR12","doi-asserted-by":"publisher","first-page":"1283","DOI":"10.1016\/S0960-9822(01)00396-7","volume":"11","author":"M Zervas","year":"2001","unstructured":"Zervas M, Somers KL, Thrall MA, Walkley SU: Critical role for glycosphingolipids in Niemann\u2013Pick disease type C. Curr Biol. 2001, 11: 1283-1287. 10.1016\/S0960-9822(01)00396-7.","journal-title":"Curr Biol"},{"key":"559_CR13","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/j.pharmthera.2009.06.003","volume":"124","author":"AP Beltrami","year":"2009","unstructured":"Beltrami AP, Cesselli D, Beltrami CA: Pluripotency rush! Molecular cues for pluripotency, genetic reprogramming of adult stem cells, and widely multipotent adult cells. Pharmacol Ther. 2009, 124: 23-30. 10.1016\/j.pharmthera.2009.06.003.","journal-title":"Pharmacol Ther"},{"key":"559_CR14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.2478\/v10039-012-0020-z","volume":"57","author":"MZ Ratajczak","year":"2012","unstructured":"Ratajczak MZ, Zuba-Surma E, Kucia M, Poniewierska A, Suszynska M, Ratajczak J: Pluripotent and multipotent stem cells in adult tissues. Adv Med Sci. 2012, 57: 1-17.","journal-title":"Adv Med Sci"},{"key":"559_CR15","doi-asserted-by":"publisher","first-page":"1294","DOI":"10.1634\/stemcells.2005-0342","volume":"24","author":"S Kern","year":"2006","unstructured":"Kern S, Eichler H, Stoeve J, Kl\u00fcter H, Bieback K: Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006, 24: 1294-1301. 10.1634\/stemcells.2005-0342.","journal-title":"Stem Cells"},{"key":"559_CR16","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1089\/scd.2007.0197","volume":"17","author":"S Anghileri","year":"2008","unstructured":"Anghileri S, Marconi S, Pignatelli A, Cifelli P, Gali\u00e9 M, Sbarbati A, Krampera M, Belluzzi O, Bonetti B: Neuronal differentiation potential of human adipose-derived mesenchymal stem cells. Stem Cells Dev. 2008, 17: 909-916. 10.1089\/scd.2007.0197.","journal-title":"Stem Cells Dev"},{"key":"559_CR17","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1016\/j.bbrc.2004.12.158","volume":"328","author":"MJ Seo","year":"2005","unstructured":"Seo MJ, Suh SY, Bae YC, Jung JS: Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun. 2005, 328: 258-264. 10.1016\/j.bbrc.2004.12.158.","journal-title":"Biochem Biophys Res Commun"},{"key":"559_CR18","doi-asserted-by":"publisher","first-page":"224","DOI":"10.1016\/j.exphem.2007.09.003","volume":"36","author":"J Case","year":"2008","unstructured":"Case J, Horvath TL, Ballas CB, March KL, Srour EF: In vitro clonal analysis of murine pluripotent stem cells isolated from skeletal muscle and adipose stromal cells. Exp Hematol. 2008, 36: 224-234. 10.1016\/j.exphem.2007.09.003.","journal-title":"Exp Hematol"},{"key":"559_CR19","doi-asserted-by":"publisher","first-page":"3438","DOI":"10.1182\/blood-2006-11-055566","volume":"110","author":"AP Beltrami","year":"2007","unstructured":"Beltrami AP, Cesselli D, Bergamin N, Marcon P, Rigo S, Puppato E, D'Aurizio F, Verardo R, Piazza S, Pignatelli A, Poz A, Baccarani U, Damiani D, Fanin R, Mariuzzi L, Finato N, Masolini P, Burelli S, Belluzzi O, Schneider C, Beltrami CA: Multipotent cells can be generated in vitro from several adult human organs (heart, liver and bone marrow). Blood. 2007, 110: 3438-3446. 10.1182\/blood-2006-11-055566.","journal-title":"Blood"},{"key":"559_CR20","doi-asserted-by":"publisher","first-page":"1225","DOI":"10.1161\/CIRCRESAHA.109.195859","volume":"104","author":"D Cesselli","year":"2009","unstructured":"Cesselli D, Beltrami AP, Rigo S, Bergamin N, D'Aurizio F, Verardo R, Piazza S, Klaric E, Fanin R, Toffoletto B, Marzinotto S, Mariuzzi L, Finato N, Pandolfi M, Leri A, Schneider C, Beltrami CA, Anversa P: Multipotent progenitor cells are present in human peripheral blood. Circ Res. 2009, 104: 1225-1234. 10.1161\/CIRCRESAHA.109.195859.","journal-title":"Circ Res"},{"key":"559_CR21","doi-asserted-by":"publisher","first-page":"8022","DOI":"10.1073\/pnas.85.21.8022","volume":"85","author":"EJ Blanchette-Mackie","year":"1988","unstructured":"Blanchette-Mackie EJ, Dwyer NK, Amende LM, Kruth HS, Butler JD, Sokol J, Comly ME, Vanier MT, August JT, Brady RO: Type-C Niemann-Pick disease: low density lipoprotein uptake is associated with premature cholesterol accumulation in the Golgi complex and excessive cholesterol storage in lysosomes. Proc Natl Acad Sci U S A. 1988, 85: 8022-8026. 10.1073\/pnas.85.21.8022.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"559_CR22","doi-asserted-by":"publisher","first-page":"890","DOI":"10.1016\/j.ajpath.2011.04.017","volume":"179","author":"S Zhou","year":"2011","unstructured":"Zhou S, Davidson C, McGlynn R, Stephney G, Dobrenis K, Vanier MT, Walkley SU: Endosomal\/lysosomal processing of gangliosides affects neuronal cholesterol sequestration in Niemann-Pick disease type C. Am J Pathol. 2011, 179: 890-902. 10.1016\/j.ajpath.2011.04.017.","journal-title":"Am J Pathol"},{"key":"559_CR23","doi-asserted-by":"publisher","first-page":"522","DOI":"10.1016\/j.copbio.2009.10.004","volume":"20","author":"DP Hunt","year":"2009","unstructured":"Hunt DP, Jahoda C, Chandran S: Multipotent skin-derived precursors: from biology to clinical translation. Curr Opin Biotechnol. 2009, 20: 522-530. 10.1016\/j.copbio.2009.10.004.","journal-title":"Curr Opin Biotechnol"},{"key":"559_CR24","doi-asserted-by":"publisher","first-page":"1082","DOI":"10.1038\/ncb1181","volume":"6","author":"KJ Fernandes","year":"2004","unstructured":"Fernandes KJ, McKenzie IA, Mill P, Smith KM, Akhavan M, Barnab\u00e9-Heider F, Biernaskie J, Junek A, Kobayashi NR, Toma JG, Kaplan DR, Labosky PA, Rafuse V, Hui CC, Miller FD: A dermal niche for multipotent adult skin-derived precursor cells. Nat Cell Biol. 2004, 6: 1082-1093. 10.1038\/ncb1181.","journal-title":"Nat Cell Biol"},{"key":"559_CR25","doi-asserted-by":"publisher","first-page":"1164","DOI":"10.1002\/stem.27","volume":"27","author":"N Gago","year":"2009","unstructured":"Gago N, Perez-Lopez V, Sanz-Jaka JP, Cormenzana P, Eizaguirre I, Bernad A, Izeta A: Age-dependent depletion of human skin-derived progenitor cells. Stem Cells. 2009, 27: 1164-1172. 10.1002\/stem.27.","journal-title":"Stem Cells"},{"key":"559_CR26","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1007\/BF00690966","volume":"66","author":"M Elleder","year":"1985","unstructured":"Elleder M, Jirasek A, Smid F, Ledvinova J, Besley GTN: Niemann\u2013Pick disease Type C: study on the nature of the cerebral storage process. Acta Neuropathol (Berl). 1985, 66: 325-336. 10.1007\/BF00690966.","journal-title":"Acta Neuropathol (Berl)"},{"key":"559_CR27","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1023\/A:1022575511354","volume":"24","author":"MT Vanier","year":"1999","unstructured":"Vanier MT: Lipid changes in Niemann-Pick disease type C brain: personal experience and review of the literature. Neurochem Res. 1999, 24: 481-489. 10.1023\/A:1022575511354.","journal-title":"Neurochem Res"},{"key":"559_CR28","doi-asserted-by":"publisher","first-page":"5886","DOI":"10.1073\/pnas.0308456101","volume":"101","author":"DE Sleat","year":"2004","unstructured":"Sleat DE, Wiseman JA, El-Banna M, Price SM, Verot L, Shen MM, Tint GS, Vanier MT, Walkley SU, Lobel P: Genetic evidence for nonredundant functional cooperativity between NPC1 and NPC2 in lipid transport. Proc Natl Acad Sci USA. 2004, 101: 5886-5891. 10.1073\/pnas.0308456101.","journal-title":"Proc Natl Acad Sci USA"},{"key":"559_CR29","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1007\/s004010050689","volume":"94","author":"PA March","year":"1997","unstructured":"March PA, Thrall MA, Brown DE, Mitchell TW, Lowenthal AC, Walkley SU: GABAergic neuroaxonal dystrophy and other cytopathological alterations in feline Niemann-Pick disease type C. Acta Neuropathol. 1997, 94: 164-172. 10.1007\/s004010050689.","journal-title":"Acta Neuropathol"},{"key":"559_CR30","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1093\/jnen\/60.1.49","volume":"60","author":"M Zervas","year":"2001","unstructured":"Zervas M, Dobrenis K, Walkley SU: Neurons in Niemann-Pick disease type C accumulate gangliosides as well as unesterified cholesterol and undergo dendritic and axonal alterations. J Neuropathol Exp Neurol. 2001, 60: 49-64.","journal-title":"J Neuropathol Exp Neurol"},{"key":"559_CR31","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1002\/cne.1149","volume":"433","author":"DC German","year":"2001","unstructured":"German DC, Quintero EM, Liang CL, Ng B, Punia S, Xie C, Dietschy JM: Selectiveneurodegeneration, without neurofibrillary tangles, in a mouse model of Niemann-Pick C disease. J Comp Neurol. 2001, 433: 415-425. 10.1002\/cne.1149.","journal-title":"J Comp Neurol"},{"key":"559_CR32","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/8904_2011_64","volume":"4","author":"L Rodr\u00edguez-Pascau","year":"2012","unstructured":"Rodr\u00edguez-Pascau L, Coll MJ, Casas J, Vilageliu L, Grinberg D: Generation of a human neuronal stable cell model for Niemann-pick C disease by RNA interference. JIMD Reports. 2012, 4: 29-37.","journal-title":"JIMD Reports"},{"key":"559_CR33","doi-asserted-by":"publisher","first-page":"2651","DOI":"10.1093\/hmg\/dds090","volume":"21","author":"MP Ordonez","year":"2012","unstructured":"Ordonez MP, Roberts EA, Kidwell C, Yuan S, Plaisted W, Goldstein LS: Disruption and therapeutic rescue of autophagy in a human neuronal model of Niemann Pick type C1. Hum Mol Genet. 2012, 21: 2651-2662. 10.1093\/hmg\/dds090.","journal-title":"Hum Mol Genet"}],"container-title":["Orphanet Journal of Rare Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1750-1172-8-34.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T19:49:08Z","timestamp":1630525748000},"score":1,"resource":{"primary":{"URL":"https:\/\/ojrd.biomedcentral.com\/articles\/10.1186\/1750-1172-8-34"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,2,21]]},"references-count":33,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,12]]}},"alternative-id":["559"],"URL":"https:\/\/doi.org\/10.1186\/1750-1172-8-34","relation":{},"ISSN":["1750-1172"],"issn-type":[{"value":"1750-1172","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,2,21]]},"assertion":[{"value":"7 September 2012","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 February 2013","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 February 2013","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"34"}}