DLK1 eta DLK2.ren karakterizazioa saguen listu guruin garapenean

103 p.; 102 p.Organogenesia seinalizazio bide ezberdinez gidatutako prozesu konplexua da. Askotan organu ezberdinen garapena seinalizazio faktore berdinez dago erregulatuta. Gainera, seinalizazio bideen ikerketa garapena ezagutarazteko erabiltzeaz aparte, minbizia eta ehunen birsorkuntza zelan gertatzen diren jakiteko ere balio digu.Lan honetan, listu-guruinaren garapena aztertu dugu. Listu-guruinaren organogenesia adrakatze morfogenesia deituriko prozesuaren bidez ematen da. Garapen prozesu hau beste organo batzuetan ere ematen da, esaterako, birikietan, giltzurrunetan eta ugatz guruinetan.NOTCH seinale bidea, garapenean dauden ehunetan, seinalerik aztertuenetako bat da, non orain dela gutxi deskribatu diren DLK1 eta DLK2 lotugai ez kanonikoak dauden. Gure lanaren helbururik nagusiena, DLK1en eta DLK2ren funtzioa ezagutzea izan zen. Batetik NOTCH seinalizazioan duten efektuak argituz eta bestetik baraila azpiko listu-guruinaren (SMG: submandibular salivary gland) garapenean jokatzen duen rola aztertuz. Hau egiteko, sagu enbrioien SMGen hazkunde organotipikoak egin genituen, sDLK1 edo DAPTrekin ( ¿-sekretasaren inibitzaile erreaktiboa). Bestalde, Dlk1 (-/-) saguarekin ere esperimentuak egin genituen. Gure emaitzek DLK1 eta DLK2 lotugaiak garapeneko listu-guruinaren maila altuetan adierazten direla erakutsi zuten. Aldiz, sagu helduko listu-guruinetan DLK1-2ren adierazpen mailak murriztu egiten dira, emaitza hauek DLK1-2 SMGaren morfogenesian parte hartu dezaketelaren ideia eman zigun. Bestalde, enbrioien SMG hazkuntzetan, sDLK1 edo DAPTrekin inhibituriko NOTCH seinale bideak, listu-guruinaren adarkatze morfogenesia kaltetzeaz aparte, inerbazioa murriztu eta azinoen barneko zelula progenitore epitelialen apoptosia eragiten zuen. NOTCH seinalearen inhibitzaileek SMGaren inerbazioa oztopatzen dutenez, karbakola (CCh) erabili genuen, inerbazio kolinergikoaren ordezko gisa bezala, modu honetan SMGaren garapena berreskuratzeko. CChak SMGaren adarkadura morfogenesia partzialki laguntzen zuela ikusi genuen, baina NOTCH seinale bidearen inhibizioa eteten zenean baino ez. Eraberean, konturatu ginen NOTCH inhibizioak sortutako morfogenesi akasduna ez zela bakarrik inerbazioa kaltetuaren ondorioz, isolatutako epitelioen morfogenesian DLK1ek eta DAPTk eragina zutela ikusi baigenuen.Dlk1 (-/-) saguaren listu-guruinen analisiarekin, DLK1en gabeziak listu-guruinaren tamaina murrizten duela ondorioztatu genuen. Gainera, pilokarpina bidezko estimulazioaren ondoren denboran neurtutako listu jariakina murriztuta dago sagu KOean. Sagu DLK1 KOaren listu-guruinen histologia eta TEMko argazkiek SMG normalak eta osasuntsuak aurkeztu zituzten, baina ama/progenitore zelulen populazioa handituta agertzen zen.Orokorrean, lan honek NOTCH seinalizazioko lotugai ez kanonikoek, DLK1 eta DLK2, saguen listu-guruinen garapenean duten garrantzia deskribatzen du. Ondorio gisa, DLK1en adierazpen mailak garapenean dauden guruinen ama/progenitore zelulen sorkuntzaren sustapen eta eragozpen arteko mekanismoaren oreka finean parte hartzen du. THESIS ABSTRACT:The organogenesis is a complex process orchestrated by different signaling pathways. The development of different organs is often regulated by the same signaling factors. The study of signaling pathways is not only to understand how organs develop, but also as a clue to investigate cancer and tissue regeneration, since these factors ale also implicated in these processes. In this work, we studied the salivary gland development. The salivary gland organogenesis occurs by a process called branching morphogenesis. This process also occurs in other organs such as mammary gland, lung and kidney. One of the most studied signaling pathways in organogenesis of developing tissues is NOTCH, where the non-canonical ligands DLK1 and DLK2 have been recently described. The main goal of our work was to elucidate the function of DLK1 and DLK2 in terms of NOTCH signaling and their role in the submandibular salivary gland (SMG) development. For this purpose, we used an in vitro organotypic culture of SMG rudiments with sDLK1 or DAPT (inhibitor reagent of ¿-secretase). For our study, we also performed experiments with null-Dlk1 mice. Our results demonstrate that these ligands are highly expressed in the development of the mouse SMG and function as NOTCH signaling inhibitors. In adult mice, DLK1-2 expression decrease, what make us think the possible role of DLK1-2 in the morphogenesis of the SMG. In SMG cultures, the inhibition of NOTCH signaling pathway, by either sDLK1 or DAPT, reduced SMG branching morphogenesis, impaired innervation and produced apoptosis in the inner epithelial progenitor cells of the developing end buds. As inhibitors of NOTCH disrupted SMG innervation, we employed a cholinergic activation reagent, carbachol (CCh), to rescue the development of the SMG. We found that CCh managed to partially recover the branching of the SMG, but only when the inhibitory effect ceased. In this context, we realized that the impaired SMG branching morphogenesis by NOTCH signaling inhibition was not only a consequence of the reduced innervation, but a DLK1 or DAPT direct effect on the SMG epithelia morphogenesis.From the analysis of the Dlk1 (-/-) mice, we concluded that DLK1 absence results in a reduced salivary gland size. As a consequence these mice produced less saliva in time after pilocarpine stimulation. DLK1-KO mice SMG histology and transmission electron microscopy showed a healthy and normal SMG, although the number of epithelial stem/progenitor cells amount increased.In overall, this work describes the importance and mechanisms of function of NOTCH non-canonical ligands DLK1 and DLK2 in the development of the salivary gland. In conclusion, we propose that the levels of DLK1 in the normal development of the salivary gland need a fine balance between the control mechanisms of inhibiting and promoting stem cells

Minbiziaren aurkako CAR-T zelulen bidezko immunoterapia

The last years immunotherapy has become a useful strategy in the fight against cancer. One of the most popular therapies is called CAR-T. In this treatment, the patient’s T lymphocytes are removed and modified in the laboratory to enhance the action against the tumour by placing a type of “sensor” on its surface (CAR receptor). Thus, the patient is re- introduced with the transformed lymphocytes, which search for, recognize, and attack tumour cells. In this review, we will explain the structure, mode of generation, and adverse effects of CAR receptors on therapy. Finally, CAR-T immunotherapy has been shown to be effective in several blood tumours, as well as in certain types of solid tumors.; Azkenengo urteotan immunoterapia minbiziari aurre egiteko estrategia baliagarria bihurtu da. Terapia ezagunenetako bati CAR-T deritzo. Tratamendu honetan, gaixoaren T linfozitoak atera eta laborategian aldatu egiten dira tumorearen aurkako ekintza hobetzeko, "sentsore" mota bat jarrita haren gainazalean (CAR errezeptorea). Horrela, pazienteari berriro linfozito eraldatuak sartzen zaizkio, hauek tumore-zelulak bilatu, ezagutu eta erasotzeko. Berrikuspen honetan, CAR errezeptoreen egitura, sortzeko era eta terapiaren efektu desiragaitzak azalduko ditugu. Azkenik, CAR-T immunoterapia odoleko hainbat tumoretan eraginkorra dela frogatu da, bai eta tumore solido mota batzuetan ere

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

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)

GeromiRs Are Downregulated in the Tumor Microenvironment during Colon Cancer Colonization of the Liver in a Murine Metastasis Model

Cancer is a phenomenon broadly related to ageing in various ways such as cell cycle deregulation, metabolic defects or telomerases dysfunction as principal processes. Although the tumor cell is the main actor in cancer progression, it is not the only element of the disease. Cells and the matrix surrounding the tumor, called the tumor microenvironment (TME), play key roles in cancer progression. Phenotypic changes of the TME are indispensable for disease progression and a few of these transformations are produced by epigenetic changes including miRNA dysregulation. In this study, we found that a specific group of miRNAs in the liver TME produced by colon cancer called geromiRs, which are miRNAs related to the ageing process, are significantly downregulated. The three principal cell types involved in the liver TME, namely, liver sinusoidal endothelial cells, hepatic stellate (Ito) cells and Kupffer cells, were isolated from a murine hepatic metastasis model, and the miRNA and gene expression profiles were studied. From the 115 geromiRs and their associated hallmarks of aging, which we compiled from the literature, 75 were represented in the used microarrays, 26 out of them were downregulated in the TME cells during colon cancer colonization of the liver, and none of them were upregulated. The histone modification hallmark of the downregulated geromiRs is significantly enriched with the geromiRs miR-15a, miR-16, miR-26a, miR-29a, miR-29b and miR-29c. We built a network of all of the geromiRs downregulated in the TME cells and their gene targets from the MirTarBase database, and we analyzed the expression of these geromiR gene targets in the TME. We found that Cercam and Spsb4, identified as prognostic markers in a few cancer types, are associated with downregulated geromiRs and are upregulated in the TME cells.This work was supported by grants from Instituto de Salud Carlos III (AC17/00012), cofounded by the European Union projects (European Regional Development Fund/European Science Foundation, Investing in your future), (ERA-Net program EracoSysMed, JTC-2 2017) and (H2020-FETOPEN, Circular Vision, Project 899417); Diputación Foral de Gipuzkoa and the Department of Economic Development and Infrastructures of the Basque Government (DFG109/20) and the Department of Economic Development and Infrastructures of the Basque Government (DFG109/Grants Health Department of the Basque Government (Spain), RIS3 call, Exp. No. 2020333039 and 2020333001. 20)

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

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

Wnt-3a Induces Epigenetic Remodeling in Human Dental Pulp Stem Cells

Dental pulp stem cells (DPSCs) from adult teeth show the expression of a very complete repertoire of stem pluripotency core factors and a high plasticity for cell reprogramming. Canonical Wnt and Notch signaling pathways regulate stemness and the expression of pluripotency core factors in DPSCs, and even very short-term (48 h) activations of the Wnt pathway induce a profound remodeling of DPSCs at the physiologic and metabolic levels. In this work, DPSC cultures were exposed to treatments modulating Notch and Wnt signaling, and also induced to differentiate to osteo/adipocytes. DNA methylation, histone acetylation, histone methylation, and core factor expression levels where assessed by mass spectroscopy, Western blot, and qPCR. A short-term activation of Wnt signaling by WNT-3A induced a genomic DNA demethylation, and increased histone acetylation and histone methylation in DPSCs. The efficiency of cell reprogramming methods relies on the ability to surpass the epigenetic barrier, which determines cell lineage specificity. This study brings important information about the regulation of the epigenetic barrier by Wnt signaling in DPSCs, which could contribute to the development of safer and less aggressive reprogramming methodologies with a view to cell therapy.This work was funded by the UPV/EHU (GIU16/66, UFI 11/44; to F.U.), the Basque Government (GV/EJ; Ikerketa Taldeak IT831-13; to G.I. and ELKARTEK KK-2019-00093; to F.U.) and ISCIII (DTS18/00142; to N.S.)

Role of Furin in Colon Cancer Stem Cells Malignant Phenotype and Expression of LGR5 and NANOG in KRAS and BRAF-Mutated Colon Tumors

Proprotein convertases or PCs are known to regulate the malignant phenotype of colon cancer cells by different mechanisms, but their effects on cancer stem cells (CSCs) have been less widely investigated. Here, we report that PCs expression is altered in colon CSCs, and the inhibition of their activity reduced colon CSCs growth, survival, and invasion in three-dimensional spheroid cultures. In vivo, repression of PCs activity by the general PC inhibitors α1-PDX, Spn4A, or decanoyl-RVKR-chloromethylketone (CMK) significantly reduced tumor expression levels of the stem cell markers LGR5 and NANOG that are associated with reduced tumor xenografts. Further analysis revealed that reduced tumor growth mediated by specific silencing of the convertase Furin in KRAS or BRAF mutated-induced colon tumors was associated with reduced expression of LGR5 and NANOG compared to wild-type KRAS and BRAF tumors. Analysis of various calcium regulator molecules revealed that while the calcium-transporting ATPase 4 (ATP2B4) is downregulated in all the Furin-silenced colon cancer cells, the Ca2+-mobilizing P2Y receptors, was specifically repressed in BRAF mutated cells and ORAI1 and CACNA1H in KRAS mutated cells. Taken together, our findings indicate that PCs play an important role in the malignant phenotype of colon CSCs and stem cell markers’ expression and highlight PCs repression, particularly of Furin, to target colon tumors with KRAS or BRAF mutation.This research was funded by La Region Nouvelle Aquitaine, Siric Brio, Ligue Contre le Cancer, Planete Vegetal and INSERM. M.J.A.-B. was supported by a grant PID2020-119715GB-I00 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”, and by the European Union H2020-FETOPEN Project 899417 Circular Vision

An Integrative Omics Approach Reveals Involvement of BRCA1 in Hepatic Metastatic Progression of Colorectal Cancer

(1) Background & Aims: The roles of different cells in the tumor microenvironment (TME) are critical to the metastatic process. The phenotypic transformation of the liver cells is one of the most important stages of the hepatic metastasis progression of colorectal cancer (CRC). Our aim was to identify the major molecules (i.e., genes, miRNAs and proteins) involved in this process. (2) Methods: We isolated and performed whole-genome analysis of gene, miRNA, and protein expression in three types of liver cells (Ito cells, Kupffer cells, and liver sinusoidal endothelial cells) from the TME of a murine model of CRC liver metastasis. We selected the statistically significant differentially expressed molecules using the Student’s t-test with Benjamini-Hochberg correction and performed functional statistically-significant enrichment analysis of differentially expressed molecules with hypergeometric distribution using the curated collection of molecular signatures, MSigDB. To build a gene-miRNA-protein network centered in Brca1, we developed a software package (miRDiana) that collects miRNA targets from the union of the TargetScan, MicroCosm, mirTarBase, and miRWalk databases. This was used to search for miRNAs targeting Brca1. We validated the most relevant miRNAs with real-time quantitative PCR. To investigate BRCA1 protein expression, we built tissue microarrays (TMAs) from hepatic metastases of 34 CRC patients. (3) Results: Using integrated omics analyses, we observed that the Brca1 gene is among the twenty transcripts simultaneously up-regulated in all three types of TME liver cells during metastasis. Further analysis revealed that Brca1 is the last BRCA1-associated genome surveillance complex (BASC) gene activated in the TME. We confirmed this finding in human reanalyzing transcriptomics datasets from 184 patients from non-tumor colorectal tissue, primary colorectal tumor and colorectal liver metastasis of the GEO database. We found that the most probable sequence of cell activation during metastasis is Endothelial→Ito→Kupffer. Immunohistochemical analysis of human liver metastases showed the BRCA1 protein was co-localized in Ito, Kupffer, and endothelial cells in 81.8% of early or synchronous metastases. However, in the greater part of the metachronous liver metastases, this protein was not expressed in any of these TME cells. (4) Conclusions: These results suggest a possible role of the co-expression of BRCA1 in Ito, Kupffer, and sinusoidal endothelial cells in the early occurrence of CRC liver metastases, and point to BRCA1 as a potential TME biomarker.D.G. and M.J.A.-B. have been supported by Grants DFG113/18 from Diputación Foral de Gipuzkoa (DFG), Spain, Ministry of Economy and Competitiveness, Spain, MINECO Grant BFU2016-77987-P and Instituto de Salud Carlos III (AC17/00012) Grant co-funded by the European Union (Eracosysmed/H2020 Grant Agreement No. 643271) and European Union (H2020-FETOPEN, Project 899417). D.G., M.J.A.-B. and I.B. have been supported by Grants Health Department of the Basque Government (Spain), RIS3 call, Exp. No. 2020333039 and 2020333001

Osteogenic differentiation of human dental pulp stem cells in decellularised adipose tissue solid foams

3D cell culture systems based on biological scaffold materials obtainable from both animal and human tissues constitute very interesting tools for cell therapy and personalised medicine applications. The white adipose tissue (AT) extracellular matrix (ECM) is a very promising biomaterial for tissue engineering due to its easy accessibility, malleability and proven biological activity. In the present study, human dental pulp stem cells (hDPSCs) were combined in vitro with ECM scaffolds from porcine and human decellularised adipose tissues (pDAT, hDAT) processed as 3D solid foams, to investigate their effects on the osteogenic differentiation capacity and bone matrix production of hDPSCs, compared to single-protein-based 3D solid foams of collagen type I and conventional 2D tissue-culture-treated polystyrene plates. pDAT solid foams supported the osteogenic differentiation of hDPSCs to similar levels to collagen type I, as assessed by alkaline phosphatase and alizarin red stainings, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and osteocalcin/bone gamma-carboxyglutamate protein (BGLAP) immunostaining. Interestingly, hDAT solid foams showed a markedly lower capacity to sustain hDPSC osteogenic differentiation and matrix calcification and a higher capacity to support adipogenesis, as assessed by RT-qPCR and oil red O staining. White ATs from both human and porcine origins are relatively abundant and available sources of raw material to obtain high quality ECM-derived biomedical products. These biomaterials could have promising applications in tissue engineering and personalised clinical therapy for the healing and regeneration of lesions involving not only a loss of calcified bone but also its associated soft non-calcified tissues.This research was supported by the Basque Government (ELKARTEK program PLAKA KK- 2019-00093; to NB), MICINN retos I+D+i (PID2019- 104766RB-C21, to JRP) and UPV/EHU (PPGA20/22; to FU, GI). The authors would like to thank the staff members of the SGIKER services of the UPV/EHU: Lipidomic service (Beatriz Abad) and Analytical Microscopy (Ricardo Andrade, Alejandro Díez-Torre and Irene Fernández) for their technical assistance

BDNF and NT3 Reprogram Human Ectomesenchymal Dental Pulp Stem Cells to Neurogenic and Gliogenic Neural Crest Progenitors Cultured in Serum-Free Medium

Background/Aims: Human Dental Pulp Stem Cells (hDPSCs) are one of the most promising types of cells to regenerate nerve tissues. Standard DMEM+10% fetal bovine serum (FBS) culture medium allows a fast expansion of hDPSC as a surface-adherent cell monolayer. However, the use of FBS also compromises the clinical use of these protocols, and its long-term presence favors hDPSCs differentiation toward mesenchymal cell-derived lineages, at the expense of a reduced capability to generate neural cells. The objective of this work was to characterize the role of neurotrophin signaling on hDPSCs using a serum-free culture protocol, and to assess the neurogenic and gliogenic capacity of hDPSCs for future nerve tissue bioengineering and regeneration. Methods: We compared the different expression of neurotrophin receptors by RT-PCR, Q-PCR, and IF of hDPSCs cultured with different growth media in the presence or absence of serum. Moreover, we assessed the response of hDPSCs to stimulation of neurotransmitter receptors by live cell calcium imaging under these different media. Finally, we compared the osteogenic potential of hDPSCs by Alizarin red staining, and the differentiation to gliogenic/neurogenic fates by immunostaining for Schwann lineage and neuronal lineage markers. We tested a commercial serum-free medium designed for the growth of mesenchymal stem cells: StemPro MSCTM (STP). Results: hDPSCs cultured in STP generated small non-adherent floating dentospheres that showed very low proliferation rates, in contrast to standard FBS-containing medium. We found that hDPSCs grown in STP conditions overexpressed neurotrophin receptor genes NTRK2 (TrkB) and NTRK3 (TrkC). Interestingly, the stimulation of these receptors by adding their respective ligands BDNF and NT-3 to STP medium enhanced the neural crest (NC) progenitor features of cultured hDPSCs. We observed a 10 to 100-fold increase of migratory NC cell markers HNK1 and P75NTR, and a significant overexpression of pluripotency core factors SOX2, OCT4 and NANOG. Moreover, hDPSCs cultured in BDNF/NT-3 supplemented STP showed a largely increased potential to differentiate towards neuronal and Schwann glial lineage cells, assessed by positive immunostaining for DCX, NeuN and S100ß, p75NTR markers, respectively. Conclusion: Our results demonstrate that the use of BDNF and NT-3 combined with STP induced the partial reprogramming of ectomesenchymal hDPSCs to generate early NC progenitor cells, which are far more competent for neuronal and glial differentiation than hDPSCs grown in the presence of FBS