12 research outputs found

    Acoustophoretic trapping of particles by bubbles in microfluidics

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    We present in this paper a simple method to produce strategic acoustic particle capture sites in microfluidic channels in a controlled way. Air bubbles are intermittently injected into a micro-capillary with rectangular cross section during a flow motion of liquid suspensions containing micron-sized particles or particles to create bubble-defined “micro-gaps” with size close to 200 µm and spheroidal geometry. The establishment of a 3D standing acoustic wave inside the capillary at a frequency close to 1 MHz produces different radiation forces on solid particles and bubbles, and acoustic streaming around the bubble. While the sample flows, part of the particles collect along the acoustic pressure node established near the central axis and continue circulating aligned through the capillary until reaching the end, where are released enriched. Meanwhile, the bubble travels very fast toward positions of maximum pressure amplitude beside the channel wall, driven by Bjerkness forces, and attach to it, remaining immovable during the acoustic actuation. Some particles adhere to its membrane trapped by the acoustic streaming generated around the oscillating bubble. Changes of frequency were applied to analyze the influence of this parameter on the bubble dynamics, which shows a complete stability once attached to the channel wall. Only increasing the flow motion induces the bubble displacements. Once reached the open air at the end of the capillary, the bubble disappears releasing the trapped particles separated from their initial host suspension with a purity degree. The device presents a very simple geometry and a low-cost fabrication.This work is financed by the Spanish National Plan project PID2021-128985OB-I00 funded by the Spanish Ministery of Science and Innovation MICINN and CSIC-Intramural project

    Study of neurogenic functional potential of dental pulp stem cells (DPSCs) in neural regeneration

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    El capítulo II (Results and Discussion) está sujeto a confidencialidad por el autor. 112 p.Ezaguna da hortz muin helduak zelula ama populazio garrantzitsu bat duela barnean, hauei, hortz muineko zelula amak deritze (ingelesez, dental pulp stem cells, DPSCs). Oinarrizko egoeran aurkezten dituzten zelula neuralen markatzaileak, zelula ama neuralek baino irisgarritasun handiagoa, jatorrizko organoaz kanpo hedatzeko ahalmena eta neurodesberdintzapen ahalmenak, bakarrak bilakatzen dituzte giza DPSCs-ak (gDPSCs-ak) gorputz helduan aurkitu daitezkeen beste zelulekiko. Izan ere, birsorkuntza neuraleko terapia zelular autologoetan erabilgarriak izan daitezke. Normalean erabiltzen diren hazkuntza eta mantentze medioek behi serum fetala (BFS) dute konposatu garrantzitsuena bezala, zeinek gDPSCs-en hazkuntza azkarra eta azalerako geruza bakar itxurako hazkuntza zelularra eragiten duen. Era berean, BFS-aren presentziak alergiak, zein erreakzio immunearen aktibazioa eragin ditzakela deskribatua dagoin vivo, konposatu honen erabilera tratamendu klinikoetatik baztertzea ezinbestekoa delarik. BFS-ak sor ditzakeen arazo medikotik at erabilgarritasun arazoak ere sor ditzake, izan ere, serumaren presentzian gDPSCs-ak leinu mesenkimaletara bideratzen direla, eta beraz, potentzial neurogenikoa galera dagoela aztertu dute zenbait autorek literaturan. BFS-aren erabilerak dakartzan eragozpen guztiek ezinbestekoa egiten dute serum gabeko eragile neurogeniko berritzaileen diseinua eta hobekuntza.Hurrengo lanean, serum gabeko hazkuntza mediotan ereindako zelulen ezaugarriak ezagutzea izan da gure xedea. Lortutako emaitzek inolako aldaketa genetiko edo epigenetikorik jasan izan ez duten gDPSCs-ak, zelula ama neuralek (ingelesez, neural stem cells, NSCs) sortzen dituzten neuro-esfera egituren antzeko hortz-esferak sortzeko duten ahalmena berretsi zuten. Hortaz aparte, bai serum gabeko zein serum-dun hazkuntza mediotan hazitako gDPSCs-ek zelula ama mesenkimal izaera eta ezaugarri pluripotentzialak mantentzeko gai zirela frogatu genuen. Hala ere, erabili genituen serum gabeko hazkuntza ingurune espezifikoek ezaugarri desberdinak erantsi zizkien gDPSCs-ei. gDPSCs-ak Neurocult¿ desberdintzapen neurogeniko medioan, NSCs-ak egunerokotasunez desberdintzeko erabiltzen den serum gabeko hazkuntza medioa, izan zirenean haziak, hauek glia eta neurona markatzaile helduak adierazteko gaitasuna zutela ikusi genuen. Honetaz aparte, markatzaile ama mesenkimal eta neural goiztiarrak adierazten dituzten genetikoki aldatu bariko gDPSCs-ez gain, CD31, markatzaile endoteliala, adierazten zuten gDPSCs-ak deskribatzen lehenak izan gara serum gabeko hazkuntza ingurunea erabiliz. Kasu honetan, Neurocult¿ hedapenerako medioa erabili genuen. Gainera, timo-gabeko xagu biluzietan garezur barneko gDPSCs-en txertaketa eta hilabete baten ostera, CD31+, Nestina + zelulak burmuineko odol-hodien zelula endotelialekin batera elkarbizitzen eta hauen laminina adierazpena areagotzen aurkitu ziren. Bestalde, gDPSC-ak StemPro MSCTM (STP) izeneko, zelula ama mesenkimalentzako hazkuntza ingurune komertzialean haziak izan zirenen. Era berean, STP-k zelulei eskainitako aldaketa molekular eta fisiologikoak erabiliz, markatzaile pluripotentzialen eta gandor neuralen (GN) markatzaileen handipen bat antzeman genuen gDPSCs-ek aurkeztutako ahalmen neurogenikoaren igotzearekin batera. Honek guztiak, gandor neuraleko ama zeluletarantz (ingelesez, neural crest stem cells, NCSCs), hau da leinu primitibo baterantz atzera pausu bat iradokitzen du.Gure emaitzek, serum gabeko erabilera gDPSC-ak hazteko egokia dela adierazten dute. Ez ziren bakarrik serum-aren erabilerarekin erlazionatutako arazoak ekidin, baizik eta leinu neuraletara asko erraztu dezakeen gandor neuraleko ama izaeraren agerpena lortu zen. Neurodesberdintzapenetik harago, serum gabeko eragileak erabiliz gDPSCs-tatik eratorritako zelula endotelial funtzionalak, burmuinean integratzeko eta odol hodiak sortzeko gaitasuna erakutsi zuten, lortzen lehen taldea izan gara .Laburbilduz, gDPSCs-en zelulen bideragarritasunarekin, amatasunarekin, ahalmen neurogenikoarekin zein sistema biziekin izan dezaketen elkarrekintzekin zerikusia duten serum gabeko zelula hazkuntzek sor ditzaketen zalantzak argituak izan dira. Gorago aipatutako ezaugarriekin batera gDPSCs-ek erakutsitako gaitasun neurogeniko eta berriki aztertutako zelula endotelialetara desberdintzeko ahalmenak ikusita, gDPSCs-ak nerbio-ehun birsortze terapietan erabiltzeko gizaki helduetan aurki daitezkeen zelula ama aproposenak direla ondorioztatu daiteke

    Study of neurogenic functional potential of dental pulp stem cells (DPSCs) in neural regeneration

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    El capítulo II (Results and Discussion) está sujeto a confidencialidad por el autor. 112 p.Ezaguna da hortz muin helduak zelula ama populazio garrantzitsu bat duela barnean, hauei, hortz muineko zelula amak deritze (ingelesez, dental pulp stem cells, DPSCs). Oinarrizko egoeran aurkezten dituzten zelula neuralen markatzaileak, zelula ama neuralek baino irisgarritasun handiagoa, jatorrizko organoaz kanpo hedatzeko ahalmena eta neurodesberdintzapen ahalmenak, bakarrak bilakatzen dituzte giza DPSCs-ak (gDPSCs-ak) gorputz helduan aurkitu daitezkeen beste zelulekiko. Izan ere, birsorkuntza neuraleko terapia zelular autologoetan erabilgarriak izan daitezke. Normalean erabiltzen diren hazkuntza eta mantentze medioek behi serum fetala (BFS) dute konposatu garrantzitsuena bezala, zeinek gDPSCs-en hazkuntza azkarra eta azalerako geruza bakar itxurako hazkuntza zelularra eragiten duen. Era berean, BFS-aren presentziak alergiak, zein erreakzio immunearen aktibazioa eragin ditzakela deskribatua dagoin vivo, konposatu honen erabilera tratamendu klinikoetatik baztertzea ezinbestekoa delarik. BFS-ak sor ditzakeen arazo medikotik at erabilgarritasun arazoak ere sor ditzake, izan ere, serumaren presentzian gDPSCs-ak leinu mesenkimaletara bideratzen direla, eta beraz, potentzial neurogenikoa galera dagoela aztertu dute zenbait autorek literaturan. BFS-aren erabilerak dakartzan eragozpen guztiek ezinbestekoa egiten dute serum gabeko eragile neurogeniko berritzaileen diseinua eta hobekuntza.Hurrengo lanean, serum gabeko hazkuntza mediotan ereindako zelulen ezaugarriak ezagutzea izan da gure xedea. Lortutako emaitzek inolako aldaketa genetiko edo epigenetikorik jasan izan ez duten gDPSCs-ak, zelula ama neuralek (ingelesez, neural stem cells, NSCs) sortzen dituzten neuro-esfera egituren antzeko hortz-esferak sortzeko duten ahalmena berretsi zuten. Hortaz aparte, bai serum gabeko zein serum-dun hazkuntza mediotan hazitako gDPSCs-ek zelula ama mesenkimal izaera eta ezaugarri pluripotentzialak mantentzeko gai zirela frogatu genuen. Hala ere, erabili genituen serum gabeko hazkuntza ingurune espezifikoek ezaugarri desberdinak erantsi zizkien gDPSCs-ei. gDPSCs-ak Neurocult¿ desberdintzapen neurogeniko medioan, NSCs-ak egunerokotasunez desberdintzeko erabiltzen den serum gabeko hazkuntza medioa, izan zirenean haziak, hauek glia eta neurona markatzaile helduak adierazteko gaitasuna zutela ikusi genuen. Honetaz aparte, markatzaile ama mesenkimal eta neural goiztiarrak adierazten dituzten genetikoki aldatu bariko gDPSCs-ez gain, CD31, markatzaile endoteliala, adierazten zuten gDPSCs-ak deskribatzen lehenak izan gara serum gabeko hazkuntza ingurunea erabiliz. Kasu honetan, Neurocult¿ hedapenerako medioa erabili genuen. Gainera, timo-gabeko xagu biluzietan garezur barneko gDPSCs-en txertaketa eta hilabete baten ostera, CD31+, Nestina + zelulak burmuineko odol-hodien zelula endotelialekin batera elkarbizitzen eta hauen laminina adierazpena areagotzen aurkitu ziren. Bestalde, gDPSC-ak StemPro MSCTM (STP) izeneko, zelula ama mesenkimalentzako hazkuntza ingurune komertzialean haziak izan zirenen. Era berean, STP-k zelulei eskainitako aldaketa molekular eta fisiologikoak erabiliz, markatzaile pluripotentzialen eta gandor neuralen (GN) markatzaileen handipen bat antzeman genuen gDPSCs-ek aurkeztutako ahalmen neurogenikoaren igotzearekin batera. Honek guztiak, gandor neuraleko ama zeluletarantz (ingelesez, neural crest stem cells, NCSCs), hau da leinu primitibo baterantz atzera pausu bat iradokitzen du.Gure emaitzek, serum gabeko erabilera gDPSC-ak hazteko egokia dela adierazten dute. Ez ziren bakarrik serum-aren erabilerarekin erlazionatutako arazoak ekidin, baizik eta leinu neuraletara asko erraztu dezakeen gandor neuraleko ama izaeraren agerpena lortu zen. Neurodesberdintzapenetik harago, serum gabeko eragileak erabiliz gDPSCs-tatik eratorritako zelula endotelial funtzionalak, burmuinean integratzeko eta odol hodiak sortzeko gaitasuna erakutsi zuten, lortzen lehen taldea izan gara .Laburbilduz, gDPSCs-en zelulen bideragarritasunarekin, amatasunarekin, ahalmen neurogenikoarekin zein sistema biziekin izan dezaketen elkarrekintzekin zerikusia duten serum gabeko zelula hazkuntzek sor ditzaketen zalantzak argituak izan dira. Gorago aipatutako ezaugarriekin batera gDPSCs-ek erakutsitako gaitasun neurogeniko eta berriki aztertutako zelula endotelialetara desberdintzeko ahalmenak ikusita, gDPSCs-ak nerbio-ehun birsortze terapietan erabiltzeko gizaki helduetan aurki daitezkeen zelula ama aproposenak direla ondorioztatu daiteke

    Vasculogenesis from Human Dental Pulp Stem Cells Grown in Matrigel with Fully Defined Serum-Free Culture Media

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    The generation of vasculature is one of the most important challenges in tissue engineering and regeneration. Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types to induce vasculogenesis and angiogenesis as they not only secrete vascular endothelial growth factor (VEGF) but can also differentiate in vitro into both endotheliocytes and pericytes in serum-free culture media. Moreover, hDPSCs can generate complete blood vessels containing both endothelial and mural layers in vivo, upon transplantation into the adult brain. However, many of the serum free media employed for the growth of hDPSCs contain supplements of an undisclosed composition. This generates uncertainty as to which of its precise components are necessary and which are dispensable for the vascular differentiation of hDPSCs, and also hinders the transfer of basic research findings to clinical cell therapy. In this work, we designed and tested new endothelial differentiation media with a fully defined composition using standard basal culture media supplemented with a mixture of B27, heparin and growth factors, including VEGF-A165 at different concentrations. We also optimized an in vitro Matrigel assay to characterize both the ability of hDPSCs to differentiate to vascular cells and their capacity to generate vascular tubules in 3D cultures. The description of a fully defined serum-free culture medium for the induction of vasculogenesis using human adult stem cells highlights its potential as a relevant innovation for tissue engineering applications. In conclusion, we achieved efficient vasculogenesis starting from hDPSCs using serum-free culture media with a fully defined composition, which is applicable for human cell therapy purposes.This work was financed by the «Ramón y Cajal» program RYC-2013-13450 (JRP); UPV/EHU (GIU16/66, UFI 11/44 and COLAB19/03; FU); MINECO Retos I + D + I (SAF2015-70866-R; JRP, PID2019-104766RB-C21; JRP); Basque Government (GV/EJ; IT831-13; GI) TERSAFURNA-2020333039 and ELKARTEK KK-2019-00093. J.L. has a grant from UPV/EHU (DOKBERRI 2019(DOCREC19/49)). I.I. has a grant from the Basque Government (PRE_2019_2_0300 or GIU16/66)

    Notch/Wnt Cross-Signalling Regulates Stemness Of Dental Pulp Stem Cells Through Expression Of Neural Crest And Core Pluripotency Factors

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    Dental pulp stem cells (DPSCs) from adult teeth express neural crest (NC) markers together with core transcriptional factors associated with stem cell pluripotency, such as Oct4a, Sox2, c-Myc, Rex1, Stella/Dppa3, Ssea1/Fut4, Lin28 and Nanog. The possibility to boost the natural stemness features of DPSCs by mild methods, that do not involve gene and/or chromatin modification or gene transfection, is highly desirable for cell therapy. Canonical Wnt and Notch are two highly conserved developmental signalling pathways that are involved in NC emergence and stem cell self-renewal. We determined that both pathways coordinate to regulate the expression of core pluripotency and NC factors in DPSCs. Pharmacological inhibition of the Notch pathway for 48 h, by the gamma-secretase inhibitor N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT), abolished the expression of NC and core factors. In addition, it induced a silencing of the canonical Wnt signalling and a clear reduction in the stemness potential of DPSCs, as shown by a reduced ability to generate mature, fully differentiated osteoblasts and adipocytes. Conversely, pharmacological activation of the Wnt pathway for 48 h, by either the glycogen synthase kinase 3 beta (GSK3-beta) inhibitor 6-bromoindirubin-3'-oxime (BIO) or the human recombinant protein Wnt-3a, not only largely increased the expression of NC and core factors, but also increased the efficiency of DPSCs to differentiate into mature osteoblasts and adipocytes. These results showed that a short preconditioning activation of Wnt/Notch signalling by small molecules and/or recombinant proteins enhanced the stemness and potency of DPSCs in culture, which could be useful for optimising the therapeutic use of these and other tissue-specific stem cells.Technical and human support provided by the analytical microscopy service of SGIKER (UPV/EHU, MINECO, GV/EJ, ERDF and ESF) is gratefully acknowledged. This work was funded by the UPV/EHU (GIU16/66, UFI 11/44) and the Basque Government (GV/EJ; IT831-13). V.U. received a fellowship from The Global Training Grant (GV/EJ) to fund a research stage at The Institute of Cancer Research (London, UK)

    Human DenPulp Stem Cells Grown in Neurogenic Media Differentiate Into Endothelial Cells and Promote Neovasculogenesis in the Mouse Brain

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    Dental pulp stem cells (DPSCs) have the capacity to give rise to cells with neuronal-like phenotypes, suggesting their use in brain cell therapies. In the present work, we wanted to address the phenotypic fate of adult genetically unmodified human DPSCs cultured in Neurocult (TM) (Stem Cell Technologies), a cell culture medium without serum which can be alternatively supplemented for the expansion and/or differentiation of adult neural stem cells (NSCs). Our results show that non-genetically modified human adult DPSCs cultured with Neurocult NS-A proliferation supplement generated neurosphere-like dentospheres expressing the NSC markers Nestin and glial fibrillary acidic protein (GFAP), but also the vascular endothelial cell marker CD31. Remarkably, 1 month after intracranial graft into athymic nude mice, human CD31+/CD146+ and Nestin+ DPSC-derived cells were found tightly associated with both the endothelial and pericyte layers of brain vasculature, forming full blood vessels of human origin which showed an increased laminin staining. These results are the first demonstration that DPSC-derived cells contributed to the generation of neovasculature within brain tissue, and that Neurocult and other related serum-free cell culture media may constitute a fast and efficient way to obtain endothelial cells from human DPSCs.This work was funded by "Ramon y Cajal" program RYC-2013-13450 (JRP) and RYC 2012-11137 (JME); Spanish Ministry of Economy and Competitiveness SAF2015-70866-R; UPV/EHU (GIU16/66, UFI 11/44); and Basque Government (GV/EJ; IT831-13). JL and OP-A obtained a Ph.D. fellowship from the University of the Basque Country (UPV/EHU)

    Low-intensity continuous ultrasound to inhibit cancer cell migration

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    In recent years, it has been verified that collective cell migration is a fundamental step in tumor spreading and metastatic processes. In this paper, we demonstrate for the first time how low-intensity ultrasound produces long-term inhibition of collective migration of epithelial cancer cells in wound healing processes. In particular, we show how pancreatic tumor cells, PANC-1, grown as monolayers in vitro respond to these waves at frequencies close to 1 MHz and low intensities (< 100 mW cm(-2)) for 48-72 h of culture after some minutes of a single ultrasound irradiation. This new strategy opens a new line of action to block the spread of malignant cells in cancer processes. Despite relevant spatial variations of the acoustic pressure amplitude induced in the assay, the cells behave as a whole, showing a collective dynamic response to acoustic performance. Experiments carried out with samples without previous starving showed remarkable effects of the LICUs from the first hours of culture, more prominent than those with experiments with monolayers subjected to fasting prior to the experiments. This new strategy to control cell migration demonstrating the effectiveness of LICUS on not starved cells opens a new line of action to study effects of in vivo ultrasonic actuation on tumor tissues with malignant cells. This is a proof-of-concept study to demonstrate the physical effects of ultrasound stimulation on tumor cell migration. An in-depth biological study of the effects of ultrasounds and underlying biological mechanisms is on-going but out of the scope of this article.This work is financed by the Spanish National Plan projects PID 2021-128985OB-I00: "New Non-invasive technology to inhibit growth of solid tumors by low intensity ultrasounds", DPI 2017-90147-R and intramural research project IRYCIS (2018/0240)

    Is There Such a Thing as a Genuine Cancer Stem Cell Marker? Perspectives from the Gut, the Brain and the Dental Pulp

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    The conversion of healthy stem cells into cancer stem cells (CSCs) is believed to underlie tumor relapse after surgical removal and fuel tumor growth and invasiveness. CSCs often arise from the malignant transformation of resident multipotent stem cells, which are present in most human tissues. Some organs, such as the gut and the brain, can give rise to very aggressive types of cancers, contrary to the dental pulp, which is a tissue with a very remarkable resistance to oncogenesis. In this review, we focus on the similarities and differences between gut, brain and dental pulp stem cells and their related CSCs, placing a particular emphasis on both their shared and distinctive cell markers, including the expression of pluripotency core factors. We discuss some of their similarities and differences with regard to oncogenic signaling, telomerase activity and their intrinsic propensity to degenerate to CSCs. We also explore the characteristics of the events and mutations leading to malignant transformation in each case. Importantly, healthy dental pulp stem cells (DPSCs) share a great deal of features with many of the so far reported CSC phenotypes found in malignant neoplasms. However, there exist literally no reports about the contribution of DPSCs to malignant tumors. This raises the question about the particularities of the dental pulp and what specific barriers to malignancy might be present in the case of this tissue. These notable differences warrant further research to decipher the singular properties of DPSCs that make them resistant to transformation, and to unravel new therapeutic targets to treat deadly tumors.This work has been financed by The University of The Basque Country (UPV/EHU): Grant number GIU16/66, UFI 11/44, COLAB19/03 and IKERTU-2020.0155 (to F.U), the Basque Government/Eusko Jaurkaritza: ELKARTEK KK-2019/00093 (to U.F.), and MINECO “Ramón y Cajal” program RYC-2013-13450 and MINECO PID2019-104766RB-C21 (to P.J.R.). L.J. was funded by a UPV/EHU postdoctoral fellowship DOKBERRI 2019 (DOCREC19/49) program

    Advances and Perspectives in Dental Pulp Stem Cell Based Neuroregeneration Therapies

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    Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types for regenerative therapies given their ability to grow in the absence of serum and their realistic possibility to be used in autologous grafts. In this review, we describe the particular advantages of hDPSCs for neuroregenerative cell therapies. We thoroughly discuss the knowledge about their embryonic origin and characteristics of their postnatal niche, as well as the current status of cell culture protocols to maximize their multilineage differentiation potential, highlighting some common issues when assessing neuronal differentiation fates of hDPSCs. We also review the recent progress on neuroprotective and immunomodulatory capacity of hDPSCs and their secreted extracellular vesicles, as well as their combination with scaffold materials to improve their functional integration on the injured central nervous system (CNS) and peripheral nervous system (PNS). Finally, we offer some perspectives on the current and possible future applications of hDPSCs in neuroregenerative cell therapies.This research was supported by MICINN retos I+D+i (PID2019-104766RB-C21 and RYC-2013-13450, to J.R.P.) and UPV/EHU (GIU16/66 and PPGA20/22, to F.U., G.I.; and COLAB19/03 and IKERTU-2020.0155, to F.U., J.R.S.). Y.P. was funded by a Bikaintek PhD grant from the Basque Government (20-AF-W2-2018-00001)

    Enhanced Adipogenic Differentiation of Human Dental Pulp Stem Cells in Enzymatically Decellularized Adipose Tissue Solid Foams

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    Engineered 3D human adipose tissue models and the development of physiological human 3D in vitro models to test new therapeutic compounds and advance in the study of pathophysiological mechanisms of disease is still technically challenging and expensive. To reduce costs and develop new technologies to study human adipogenesis and stem cell differentiation in a controlled in vitro system, here we report the design, characterization, and validation of extracellular matrix (ECM)-based materials of decellularized human adipose tissue (hDAT) or bovine collagen-I (bCOL-I) for 3D adipogenic stem cell culture. We aimed at recapitulating the dynamics, composition, and structure of the native ECM to optimize the adipogenic differentiation of human mesenchymal stem cells. hDAT was obtained by a two-enzymatic step decellularization protocol and post-processed by freeze-drying to produce 3D solid foams. These solid foams were employed either as pure hDAT, or combined with bCOL-I in a 3:1 proportion, to recreate a microenvironment compatible with stem cell survival and differentiation. We sought to investigate the effect of the adipogenic inductive extracellular 3D-microenvironment on human multipotent dental pulp stem cells (hDPSCs). We found that solid foams supported hDPSC viability and proliferation. Incubation of hDPSCs with adipogenic medium in hDAT-based solid foams increased the expression of mature adipocyte LPL and c/EBP gene markers as determined by RT-qPCR, with respect to bCOL-I solid foams. Moreover, hDPSC capability to differentiate towards adipocytes was assessed by PPAR-γ immunostaining and Oil-red lipid droplet staining. We found out that both hDAT and mixed 3:1 hDAT-COL-I solid foams could support adipogenesis in 3D-hDPSC stem cell cultures significantly more efficiently than solid foams of bCOL-I, opening the possibility to obtain hDAT-based solid foams with customized properties. The combination of human-derived ECM biomaterials with synthetic proteins can, thus, be envisaged to reduce fabrication costs, thus facilitating the widespread use of autologous stem cells and biomaterials for personalized medicine.This research was funded by the Basque Government (IT1751-22; to G.I.; ELKARTEK program 566 PLAKA KK-2019-00093; to N.B.), the Health Department of the Basque Government (grant No. 2021333012; to J.R.P.), and grant No. RYC-2013-13450 and grant No. PID2019-104766RB-C21 funded by MCIN/AEI/10.13039/501100011033 by the European Union (NextGenerationEU) “Plan de Recuperación Transformación y Resiliencia” (grants to J.R.P.)
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