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Possibility of Undifferentiated Human Thigh Adipose Stem Cells Differentiating into Functional Hepatocytes
Jong Hoon Lee,Kuk Han Lee,Min Ho Kim,Jun Pyo Kim
Archives of Plastic Surgery , 2012, DOI: http://dx.doi.org/10.5999/aps.2012.39.6.593
Abstract: Background This study aimed to investigate the possibility of isolating mesenchymal stemcells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stemcells (HTASCs) to differentiate into hepatocytes.Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue.Growth factors, cytokines, and hormones were added to the collagen coated dishes to inducethe undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm theexperimental results, the expression of hepatocyte-specific markers on undifferentiated anddifferentiated HTASCs was analyzed using reverse transcription polymerase chain reactionand immunocytochemical staining. Differentiation efficiency was evaluated using functionaltests such as periodic acid schiff (PAS) staining and detection of the albumin secretion levelusing enzyme-linked immunosorbent assay (ELISA).Results The majority of the undifferentiated HTASCs were changed into a more polygonalshape showing tight interactions between the cells. The differentiated HTASCs up-regulatedmRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intenselystained with anti-albumin antibody compared with undifferentiated HTASCs. PAS stainingshowed that HTASCs submitted to the hepatocyte differentiation protocol were able to morespecifically store glycogen than undifferentiated HTASCs, displaying a purple color in thecytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCscould secrete albumin, which is one of the hepatocyte markers.Conclusions MSCs were islolated from human thigh adipose tissue differentiate toheapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but alsothigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.
Alginate in cell cryopreservation processes and Its rol as a chondrogenic differentiation inducing factor
Giraldo-Hoyos Juliana María,Trochêz-Wilchez Diana Fernanda,Valencia-Serna Juliana,Zapata-Linares Natalia
Revista Ciencias Biomédicas , 2011,
Abstract: This study evaluated the influence of sodium alginate in the chondrogenic differentiationprocesses of adult stem cells, and its possible role as a non-penetrating cryopreservationagent for long periods of cell storage. In order to achieve this goal, stem cells fromadipose tissue were isolated and cultured in an alginate three-dimensional matrix andsubsequently cryopreserved during different time periods. The influence of alginate wasevaluated on the morphology of the encapsulated cells and, it was found that cells wentfrom having a fibroblastoid morphology to a rounded morphology. Additionally, afterstaining with Safranin O, proteoglycans were found in the encapsulated cells, suggestingthat this cell culture material provides the generation of a typical extracellular matrixof chondrocyte tissue. The viability of cells after being cryopreserved in the alginatematrixes during different time periods had a lower viability in most of the cases incomparison with the cells normally cryopreserved. Results suggest that sodium alginateis a good chondrogenic differentiation inducer, but not so useful for the cryopreservationprocess of adult stem cells.RESUMEN:En este estudio se evaluó la influencia que ejerce el alginato de sodio en procesos dediferenciación condrogénica de células madre adultas, así como su posible rol comoagente criopreservante no penetrante para su mantenimiento por largos periodos detiempo. Con el fin de alcanzar este objetivo, células madre obtenidas de tejido adiposo,fueron aisladas y cultivadas en una matriz tridimensional de alginato y posteriormentecriopreservadas por distintos periodos de tiempo. Se evaluó la influencia del alginatosobre la morfología de las células encapsuladas y se encontró que las células pasaronde tener una morfología fibroblastoide, a una morfología redondeada. Adicionalmente,después de la coloración con Safranina O, se encontró presencia de proteoglicanosen las células encapsuladas, lo que sugiere que el cultivo en este material, proveela generación de una matriz extracelular típica del tejido condrocítico. En cuanto a laviabilidad de las células después de ser criopreservadas en las matrices de alginato pordiferentes periodos de tiempo, tenían una menor viabilidad en la mayoría de los casos encomparación con las células normalmente criopreservadas. Dichos resultados sugierenque el alginato de sodio es un buen inductor de diferenciación condrogénica, pero no estan útil para criopreservación de células madre adultas.
In vitro differentiation of human adipose-derived mesenchymal stem cells into endothelial-like cells
Lidong Guan,Li Shaoqing,Yunfang Wang,Huimin Yue,Daqing Liu,Lijuan He,Cixian Bai,Fang Yan,Xue Nan,Shuangshuang Shi,Xuetao Pei
Chinese Science Bulletin , 2006, DOI: 10.1007/s11434-006-2055-7
Abstract: The neovascularization of ischemic tissue is a crucial initial step for the functional rehabilitation and wound healing. However, the short of seed cell candidate for the foundation of vascular network is still a big issue. Human adipose tissue derived mesenchymal stem cells (hADSCs), which possess multilineage potential, are capable of adipogenic, osteogenic, and chondrogenic differentiation. We examined whether this kind of stem cells could differentiate into endothelial-like cells and participate in blood vessel formation, and whether they could be used as an ideal cell source for therapeutic angiogenesis in ischemic diseases or vascularization of tissue constructs. The results showed that hADSCs, grown under appropriately induced conditions, displayed characteristics similar to those of vessel endothelium. The differentiated cells expressed endothelial cell markers CD34 and vWF, and had high metabolism of acetylated low-density lipoprotein and prostacyclin. In addition, the induced cells were able to form tube-like structures when cultured on matrigel. Our data indicated that induced hADSCs could exhibit characteristics of endothelial cells. Therefore, these cells, as a source of human endothelial cells, may find many applications in such realms as engineering blood vessels, endothelial cell transplantation for myocardial regeneration, and induction of angiogenesis for treatment of regional ischemia.
The use of adipose-derived stem cells for the fabrication of three-dimensional spheroids for the osteogenic differentiation
Jae-Yong Tae,Jun-Beom Park,Sung-Il Lee,Youngkyung Ko
- , 2017,
Abstract: Adipose-derived stem cells have been applied for the treatment of various diseases. This study was performed to fabricate the three-dimensional stem cell spheroids with adipose-derived stem cells and to evaluate the effects of cell number and culture duration on the viability and the osteogenic differentiation of three-dimensional cultures. Adipose-derived stem cells were purchased, and stem cell spheroids were formed using the silicon elastomer-based concave microwells in the amount of 1 × 105, 3 × 105, and 6 × 105. The morphology, the cell viability, and the osteogenic differentiation of stem cell spheroids were evaluated. Adipose-derived stem cells formed spheroids in the silicon elastomer-based concave microwells. A higher stem cell number led to spheroids with larger diameters. Most of the cells in the cell spheroids emitted green fluorescence. The relative Cell Counting Kit-8 showed that higher values were seen with higher stem cell numbers. The increase of alkaline phosphatase activity assays with longer incubation time in low dosage group and mineralized extracellular deposits were evenly noted in each group. This study clearly showed that adipose-derived stem cells formed spheroids in the silicon elastomer-based concave microwells and that osteogenic differentiation was achieved with higher activity and longer incubation time. Spheroid-based cell delivery could be used as a simple and effective strategy for stem cell therapy
Mouse adipose tissue stromal cells give rise to skeletal and cardiomyogenic cell sub-populations
Cécile Dromard,Valerie Planat-Benard
Frontiers in Cell and Developmental Biology , 2014, DOI: 10.3389/fcell.2014.00042
Abstract: We previously reported that adipose tissue could generate cardiomyocyte-like cells from crude stromal vascular fraction (SVF) in vitro that improved cardiac function in a myocardial infarction context. However, it is not clear whether these adipose-derived cardiomyogenic cells (AD-CMG) constitute a homogenous population and if AD-CMG progenitors could be isolated as a pure population from the SVF of adipose tissue. This study aims to characterize the different cell types that constitute myogenic clusters and identify the earliest AD-CMG progenitors in vitro for establishing a complete phenotype and use it to sort AD-CMG progenitors from crude SVF. Here, we report cell heterogeneity among adipose-derived clusters during their course of maturation and highlighted sub-populations that exhibit original mixed cardiac/skeletal muscle phenotypes with a progressive loss of cardiac phenotype with time in liquid culture conditions. Moreover, we completed the phenotype of AD-CMG progenitors but we failed to sort them from the SVF. We demonstrated that micro-environment is required for the maturation of myogenic phenotype by co-culture experiments. These findings bring complementary data on AD-CMG and suggest that their emergence results from in vitro events.
Cell sheet fabrication of hepatocyte-like cells differentiated from adipose tissue mesenchymal stem cells
大鼠脂肪间充质干细胞诱导为类肝细胞及其细胞片的制备
Yongbo Lu,Feng Qiu,Yongzhu Chen,Xiaojun Zhao,
卢永波,邱峰,陈咏竹,赵晓军
生物工程学报 , 2009,
Abstract: Adult pluripotent stem cells,such as mesenchymal stem cells derived from bone marrow and adipose tissue are capable of multilineage differentiation. Although autologous stem cell transplantation is an effective alternative to organ transplantation,the loss of cell viability and differentiation confinement of implanted cells has largely impaired the therapeutic efficacy. To produce biomaterial-free liver construct to integrate into living tissue,we isolated adipose mesenchymal stem cells and subjected them t...
Impact of Celosia cristata Extract on Adipogenesis of Native Human CD34+/CD31- Cells  [PDF]
Richard Fitoussi, David Estève, Anne-Sophie Delassus, Katell Vié
Journal of Cosmetics, Dermatological Sciences and Applications (JCDSA) , 2013, DOI: 10.4236/jcdsa.2013.33A2013
Abstract:

Background: The adipose tissue mainly consists of adipocytes but also contains non-adipose cells. Among them, progenitor cells represent a local pool of immature cells that, in vitro, can undergo various lineage differentiation processes. These cells are thought to contribute to normal homeostasis of the adipose tissue through adipogenesis but also to the growth of the adipose tissue under chronic energy overload. The aim of the present study is to evaluate in vitro the capacity of a Celosia cristata extract to impact the adipogenic potential of native human adipose tissue progenitor cells, i.e. commitment and differentiation towards adipogenic lineage. Methods: Native adipose tissue progenitor cells were isolated by immunoselection/depletion approaches from human subcutaneous adipose tissues. Two distinct cell culture conditions were used to assess the effect of Celosia cristata extract on commitment and differenciation of progenitor cells. Cells were cultured either in differentiation medium for 10 days in the presence/absence of Celosia cristata extracts to study the impact on differentiation or first cultured in a commitment-inducing medium, with or without Celosia cristata extract, for 48 h and then cultured 10 days in differentiation medium to assess the impact on commitment. In both experimental series, the fate of progenitor cells was studied by quantification of lipids and by

High-Fat Diets-Induced Metabolic Alterations Alter the Differentiation Potential of Adipose Tissue-Derived Stem Cells  [PDF]
Vikie Lamontagne, Souhad El Akoum, Isabelle Cloutier, Jean-Fran？ois Tanguay
Open Journal of Endocrine and Metabolic Diseases (OJEMD) , 2013, DOI: 10.4236/ojemd.2013.33027
Abstract:

Background: Adipose tissue-derived stem cells (ASC) possess the ability to differentiate into adipocytes or endothelial cells to help in the adipogenesis, vasculogenesis and vascular repair. This study aims at determining the impact of high-fat diets (HFD)-induced type 2 diabetes (T2D) on the differentiation potential of ASC. Results: C57BL/6J male mice were fed a vegetal (VD) or an animal (AD) HFD. Isolation of ACS from mice showing different levels of metabolic alterations reveals that advanced T2D did not affect the number of cells per gram of tissue. Rather, a higher proportion of inflammatory CD36+ cells was identified in HFD fed mice. Despite a marked decreased expression of adipogenic genes (aP2, C/EBPα and PPARγ2), ASC from HFD groups had a higher adipogenic potential and a lower endothelial differentiation potential in vitro compared to control. ASC from the VD group had enhanced cyclin B1 expression and had more adipogenic potential compared to AD group. Conclusion: Our results demonstrate that the metabolic modifications, linked to the nature of fatty acids in diets, modulate the differentiation potential of ASC with increased adipogenesis to the detriment of the endothelial pathway. Results highlight the importance of evaluating the ASC differentiation behavior in a context of autologous cell-based therapy for the repair of vascular tissues in diabetic patients.

Effects of melatonin on the proliferation and differentiation of rat adipose-derived stem cells
Zaminy Arash,Kashani Iraj,Barbarestani Mohammad,Hedayatpour Azim
Indian Journal of Plastic Surgery , 2008,
Abstract: Background: Osteogenesis driven by adipose-derived stem cells (ADSCs) is regulated by physiological and pathological factors. Accumulating evidence from in vitro and in vivo experiments suggests that melatonin may have an influence on bone formation. However, little is known about the effects of melatonin on osteogenesis, which thus remains to be elucidated. This study was performed to determine whether melatonin at physiological concentrations (0.01-10 nM) could affect the in vitro proliferation and osteogenic differentiation of rat ADSCs. Materials and Methods: ADSCs were isolated from the fat of adult rats. After cell expansion in culture media and through three passages, osteogenesis was induced in a monolayer culture using osteogenic medium with or without melatonin at physiological concentrations (0.01-10 nM). After four weeks, the cultures were examined for mineralization by Alizarin Red S and von Kossa staining and for alkaline phosphatase (ALP) activity using an ALP kit. Cell viability and apoptosis were also assayed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTT) assay and flow cytometry, respectively. Results: The results indicated that at physiological concentrations, melatonin suppressed proliferation and differentiation of ADSCs. These data indicate that ADSCs exposed to melatonin, had a lower ALP activity in contrast to the cells exposed to osteogenic medium alone. Similarly, mineral deposition (calcium level) also decreased in the presence of melatonin. Flow cytometry confirmed that cell growth had decreased and that the numbers of apoptotic cells had increased. Conclusion: These results suggest that the physiological concentration of melatonin has a negative effect on ADSC osteogenesis.
Differentiation of Adipose-derived Stem Cells into Schwann Cell Phenotype in Comparison with Bone Marrow Stem Cells
Zolikha Golipoor,Iraj Ragerdi Kashani,Mohammad Akbari,Gholamreza Hassanzadeh
Iranian Journal of Basic Medical Sciences , 2010,
Abstract: Objective(s)Bone marrow is the traditional source of human multipotent mesenchymal stem cells (MSCs), but adipose tissue appears to be an alternative and more readily available source. In this study, rat adipose-derived stem cells (ADSCs) were induced to differentiate into Schwann-like cells and compared with rat bone marrow stem cells (BMSCs) for their Schwann-like cells differentiation potential. Materials and MethodsBMSCs and ADSCs were characterized for expression of MSCs-specific markers, osteogenic and adipogenic differentiation. They were induced to differentiate into Schwann-like cells and analyzed for expression of the Schwann specific markers. The immunocytochemical differentiation markers were S-100 and real time quantitative Real-time polymerase chain reaction (RT-PCR) markers were S100, P75 and glial fibrillary acidic protein (GFAP). 3-(4, 5-Dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and Annexin V-Fluorescein isothiocyanate (FITC)/ Propidium iodide (PI) double labeling method were employed to detect early stage cell apoptosis.ResultsBMSCs and ADSCs showed similarities in expression of the MSC-specific markers, osteogenic and adipogenic differentiation. Both quantitative RT-PCR and immunocytochemical analysis demonstrated that BMSCs and ADSCs had equal expression of the Schwann-specific markers following Schwann-like cells differentiation. However, gene expression of P75 was higher in BMSCs compared with ADSCs. MTT assay and flow cytometry found that of the total BMSCs and ADSCs in the culture medium, 20% to 30% of the cells died, but the remaining cell population remained strongly attached to the substrate and differentiated.ConclusionComparative analysis showed that Schwann-like cell differentiation potential of ADSCs was slightly decreased in comparison with BMSCs. Therefore, BMSCs are more favorable choice than ADSCs for tissue engineering.
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