scholarly journals Elastin-Collagen Based Hydrogels as Model Scaffolds to Induce Three-Dimensional Adipocyte Culture from Adipose Derived Stem Cells

2020 ◽  
Vol 7 (3) ◽  
pp. 110 ◽  
Author(s):  
Kristen Newman ◽  
Kendra Clark ◽  
Bhuvaneswari Gurumurthy ◽  
Pallabi Pal ◽  
Amol V. Janorkar
Keyword(s):  
Stem Cells ◽  
Adipogenic Differentiation ◽  
Three Dimensional ◽  
Adipose Derived Stem Cells ◽  
Collagen Scaffolds ◽  
Collagen Concentration ◽  
Physico Chemical ◽  
Culture Models ◽  
Oil Red O Staining

This study aimed to probe the effect of formulation of scaffolds prepared using collagen and elastin-like polypeptide (ELP) and their resulting physico-chemical and mechanical properties on the adipogenic differentiation of human adipose derived stem cells (hASCs). Six different ELP-collagen scaffolds were prepared by varying the collagen concentration (2 and 6 mg/mL), ELP addition (6 mg/mL), or crosslinking of the scaffolds. FTIR spectroscopy indicated secondary bonding interactions between collagen and ELP, while scanning electron microscopy revealed a porous structure for all scaffolds. Increased collagen concentration, ELP addition, and presence of crosslinking decreased swelling ratio and increased elastic modulus and compressive strength of the scaffolds. The scaffold characteristics influenced cell morphology, wherein the hASCs seeded in the softer, non-crosslinked scaffolds displayed a spread morphology. We determined that stiffer and/or crosslinked elastin-collagen based scaffolds constricted the spreading of hASCs, leading to a spheroid morphology and yielded an enhanced adipogenic differentiation as indicated by Oil Red O staining. Overall, this study underscored the importance of spheroid morphology in adipogenic differentiation, which will allow researchers to create more physiologically-relevant three-dimensional, in vitro culture models.

scholarly journals Effect of Uniaxial Tensile Cyclic Loading Regimes on Matrix Organization and Tenogenic Differentiation of Adipose-Derived Stem Cells Encapsulated within 3D Collagen Scaffolds

2017 ◽  
Vol 2017 ◽  
pp. 1-16 ◽  
Author(s):  
Gayathri Subramanian ◽  
Alexander Stasuk ◽  
Mostafa Elsaadany ◽  
Eda Yildirim-Ayan
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Expression Profiles ◽  
Three Dimensional ◽  
Uniaxial Tensile ◽  
Adipose Derived Stem Cells ◽  
Collagen Scaffolds ◽  
Differentiation Potential ◽  
Tenogenic Differentiation ◽  
Matrix Organization

Adipose-derived mesenchymal stem cells have become a popular cell choice for tendon repair strategies due to their relative abundance, ease of isolation, and ability to differentiate into tenocytes. In this study, we investigated the solo effect of different uniaxial tensile strains and loading frequencies on the matrix directionality and tenogenic differentiation of adipose-derived stem cells encapsulated within three-dimensional collagen scaffolds. Samples loaded at 0%, 2%, 4%, and 6% strains and 0.1 Hz and 1 Hz frequencies for 2 hours/day over a 7-day period using a custom-built uniaxial tensile strain bioreactor were characterized in terms of matrix organization, cell viability, and musculoskeletal gene expression profiles. The results displayed that the collagen fibers of the loaded samples exhibited increased matrix directionality with an increase in strain values. Gene expression analyses demonstrated that ASC-encapsulated collagen scaffolds loaded at 2% strain and 0.1 Hz frequency showed significant increases in extracellular matrix genes and tenogenic differentiation markers. Importantly, no cross-differentiation potential to osteogenic, chondrogenic, and myogenic lineages was observed at 2% strain and 0.1 Hz frequency loading condition. Thus, 2% strain and 0.1 Hz frequency were identified as the appropriate mechanical loading regime to induce tenogenic differentiation of adipose-derived stem cells cultured in a three-dimensional environment.

scholarly journals Effect of the deuterium on efficiency and type of adipogenic differentiation of human adipose-derived stem cells in vitro

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Alona V. Zlatska ◽  
Roman G. Vasyliev ◽  
Inna M. Gordiienko ◽  
Anzhela E. Rodnichenko ◽  
Maria A. Morozova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipogenic Differentiation ◽  
Adipose Derived Stem Cells

scholarly journals Anti-inflammatory effect of adipose-derived stem cells in the treatment of pelvic organ prolapse

2019 ◽  
Vol 47 (12) ◽  
pp. 6303-6314
Author(s):  
Su Wang ◽  
Jian Gao ◽  
Maohuai Wang ◽  
Liquan Chen ◽  
Xiaowei Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pelvic Organ Prolapse ◽  
Adipogenic Differentiation ◽  
Pelvic Organ ◽  
Tissue Growth ◽  
Collagen I ◽  
Adipose Derived Stem Cells ◽  
Porcine Pericardium

Objective This study investigated the effect of recombinant human connective tissue growth factor (hCTGF) on rat adipose-derived stem cells (ADSCs) and explored the feasibility of using ADSCs to treat pelvic organ prolapse. Methods ADSCs were isolated from rat inguinal adipose tissue and characterized by flow cytometry and for osteogenic and adipogenic differentiation. ADSCs were treated with recombinant hCTGF and qRT-PCR was performed to detect collagen I and III expression on post-treatment days 7, 14, and 28. Osteogenic and adipogenic differentiation of ADSCs was performed to evaluate the effect of hCTGF. ADSCs were seeded in biological grafting materials, acellular porcine pericardium (APP) and acellular bovine pericardium (ABP), then implanted in the rat vagina. Histology was performed to observe inflammation among different groups. Results Collagen I and III expression in ADSCs was significantly increased, and the ability to differentiate into osteogenic and adipogenic lineages was diminished after hCTGF treatment. APP and ABP seeded with ADSCs significantly decreased inflammation and protected from degradation in vivo compared with APP and ABP only; ABP seeded with ADSCs had the lowest inflammation. Conclusion hCTGF regulates collagen I and III expression and induces ADSC differentiation in vitro. ADSCs decrease inflammation associated with APP and ABP in vivo.

scholarly journals Activating PIK3CA mutation promotes adipogenesis of adipose-derived stem cells in macrodactyly via up-regulation of E2F1

2020 ◽  
Vol 11 (7) ◽  
Author(s):  
Bin Sun ◽  
Yongkang Jiang ◽  
Hengqing Cui ◽  
Xia Fang ◽  
Gang Han ◽  
...  
Keyword(s):  
Stem Cells ◽  
Soft Tissues ◽  
Adipogenic Differentiation ◽  
Pik3ca Mutation ◽  
Adipose Derived Stem Cells ◽  
Rna Seq ◽  
Activating Mutation ◽  
Excessive Accumulation

Abstract Macrodactyly is a congenital malformation characterized by enlargement of bone and soft tissues in limbs, typically with excessive accumulation of adipose tissues. Although gain-of-function mutation of PIK3CA has been identified in macrodactyly, the mechanism of PIK3CA mutation in adipose accumulation is poorly understood. In this study, we found that adipocytes from macrodactyly were more hypertrophic than those observed in polydactyly. PIK3CA (H1047R) activating mutation and enhanced activity of PI3K/AKT pathway were detected in macrodactylous adipose-derived stem cells (Mac-ADSCs). Compared to polydactyly-derived ADSCs (Pol-ADSCs), Mac-ADSCs had higher potential in adipogenic differentiation. Knockdown of PIK3CA or inhibition by BYL-719, a potent inhibitor of PIK3CA, impaired adipogenesis of Mac-ADSCs in vitro. In vivo study, either transient treatment of ADSCs or intragastrical gavage with BYL-719 inhibited the adipose formation in patient-derived xenograft (PDX). Furthermore, RNA-seq revealed that E2F1 was up-regulated in Mac-ADSCs and its knockdown blocked the PIK3CA-promoted adipogenesis. Our findings demonstrated that PIK3CA activating mutation promoted adipogenesis of ADSCs in macrodactyly, and that this effect was exerted by the up-regulation of E2F1. This study revealed a possible mechanism for adipose accumulation in macrodactyly and suggested BYL-719 as a potential therapeutic agent for macrodactyly treatment.

scholarly journals Osteogenic Differentiation of Three-Dimensional Bioprinted Constructs Consisting of Human Adipose-Derived Stem Cells In Vitro and In Vivo

PLoS ONE ◽  
2016 ◽  
Vol 11 (6) ◽  
pp. e0157214 ◽  
Author(s):  
Xiao-Fei Wang ◽  
Yang Song ◽  
Yun-Song Liu ◽  
Yu-chun Sun ◽  
Yu-guang Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Three Dimensional ◽  
Adipose Derived Stem Cells

scholarly journals Knockdown of CDC20 Promotes Adipogenesis of Bone Marrow-derived Stem Cells

2021 ◽  
Author(s):  
Yangge Du ◽  
Yunsong Liu ◽  
Yongsheng Zhou ◽  
Ping Zhang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Tissue ◽  
Western Blot ◽  
Adipogenic Differentiation ◽  
Conditional Knockout ◽  
Qrt Pcr ◽  
Oil Red O Staining ◽  
Oil Red O

Abstract Background: Bone is a rigid organ that provides support and physical protection to vital organs of the body. Several bone loss disorders are commonly associated with increased bone marrow adipose tissue. Bone marrow mesenchymal stromal/stem cells (BMSCs) are multipotent progenitors differentiating into osteoblasts, adipocytes, and chondrocytes. CDC20 is a co-activator of APC/C, required for full ubiquitin ligase activity. In our previous study, CDC20 promoted the osteogenic commitment of BMSCs and Cdc20 conditional knockout mice suggested a decline in bone mass. In this study, we investigated the function of CDC20 in the adipogenic differentiation of BMSCs and provided a new clue between adipogenesis and osteogenesis. Methods: Lentivirus containing CDC20 shRNA was used for CDC20 knockdown in hBMSCs. Primary mBMSCs were isolated from Cdc20f/f and Sp7-Cre;Cdc20f/f mice. Adipogenesis was examined by qRT-PCR and western blot analysis of adipogenic regulators, Oil Red O staining and transplantation into nude mice. The CDC20 knockout efficiency was determined through immunochemistry, qRT-PCR and western blot of bone marrow. Accumulation of adiposity was measured through histology and staining of bone sections. Results: CDC20 expression in hBMSCs was significantly decreased during adipogenic differentiation. Knockdown of CDC20 enhanced adipogenic differentiation of hBMSCs in vitro. CDC20-knockdown hBMSCs showed more adipose tissue–like constructs in H&E staining and Oil Red O staining. Sp7-Cre;Cdc20f/f mice presented increased adipocytes in bone marrow compared with control mice. mBMSCs from Sp7-Cre;Cdc20f/f mice exerted upregulated adipogenic differentiation. Conclusions: Our findings showed that knockdown of CDC20 enhanced adipogenesis of h(m)BMSCs in vitro and in vivo. Overall, CDC20 regulated both adipogenesis and osteogenesis of BMSCs, and may lead to the development of new therapeutic target for “fatty bone” and osteoporosis.

scholarly journals In vitro Comparison of Adipogenic Differentiation in Human Adipose-Derived Stem Cells Cultured with Collagen Gel and Platelet-Rich Fibrin

2021 ◽  
Vol 54 (03) ◽  
pp. 278-283
Author(s):  
Pallavi Priyadarshini ◽  
Soumi Samuel ◽  
Basan Gowda Kurkalli ◽  
Chethan Kumar ◽  
Basavarajappa Mohana Kumar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Soft Tissue ◽  
Cell Surface ◽  
Adipogenic Differentiation ◽  
Collagen Gel ◽  
Surface Markers ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Markers ◽  
Platelet Rich Fibrin

Abstract Background: Adipose-derived stem cells (ADSCs) are the most preferred cell type, based on their phenotypic characteristics, plasticity, and favorable immunological properties for applications in soft-tissue augmentation. Hence, the present in vitro study was aimed to evaluate the adipogenic differentiation potential of human ADSCs upon culturing individually with collagen gel and platelet-rich fibrin (PRF). Materials and methods: The collected lipoaspirate was used for establishing ADSCs using enzymatic digestion method. Then, the cells were analyzed for their morphology, viability, proliferation rate, population doubling time (PDT), colony-forming ability, cell surface markers expression, and osteogenic differentiation as biological properties. Further, ADSCs were evaluated for their adipogenicity using induction media alone, and by culturing with collagen gel and PRF individually for prospective tissue augmentation. Results: ADSCs were successfully established in vitro and exhibited a fibroblast-like morphology throughout the culture period. Cells had higher viability, proliferation potential and showed their ability to form colonies. The positive expression of cell surface markers and osteogenic ability confirmed the potency of ADSCs. The ADSCs cultured on collagen gel and PRF, individually, showed higher number of differentiated adipocytes than ADSCs grown with adipogenic induction medium alone. Conclusion: The extent of lipid accumulation by ADSCs was slightly higher when cultured on collagen gel than on PRF. Additional experiments are required to confirm better suitability of scaffold materials for soft-tissue regeneration.

scholarly journals The Effect of Age on the Regenerative Potential of Human Eyelid Adipose-Derived Stem Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Mingqi Zhang ◽  
Zhuoshi Wang ◽  
Yan Zhao ◽  
Lirong Zhang ◽  
Ling Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Adipogenic Differentiation ◽  
Culture Method ◽  
Explant Culture ◽  
Adipose Derived Stem Cells ◽  
Cell Therapies ◽  
Age Related ◽  
Autologous Mesenchymal Stem Cells

Human eyelid adipose-derived stem cells (HEASCs) are a new source of autologous mesenchymal stem cells, which are derived from neuroectoderm and potentially applied in the tissue regeneration and cell therapies. Based on the prevalence of blepharoplasty in Asia and the availability of HEASCs, we investigated the effect of donor age on their characteristics and regenerative potential of HEASCs in vitro. The HEASCs were isolated from patients of three groups: (1) <20 years (n=4), (2) >20 years, <45 years (n=5), and (3) >55 years (n=4). For each group, the proliferative capacity, colony-forming ability, surface markers, differentiation ability, wound healing function, and secreted protein were contrastively evaluated and quantified for statistical analysis. It was found that HEASCs were successfully isolated and cultured by an explant culture method. The proliferative rates, osteogenic and chondrogenic differentiation potentials, wound healing ability, and the expression of TGF-β1 and fibronectin protein of HEASCs significantly decreased as age increased. However, the expression of CD90 antigen and the adipogenic differentiation showed an age-related increase in HEASCs. As many degenerative diseases increase in prevalence with age, the age-related changes of the HEASCs proliferation potential, differentiation capacity, and wound healing ability should be taken into account whenever they are intended for use in research or cytotherapy.

scholarly journals Enhanced Adipogenic Differentiation of Human Adipose-Derived Stem Cells in an in vitro Microenvironment: The Preparation of Adipose-Like Microtissues Using a Three-Dimensional Culture

Cell Medicine ◽  
2017 ◽  
Vol 9 (1-2) ◽  
pp. 35-44 ◽  
Author(s):  
Yoshitaka Miyamoto ◽  
Masashi Ikeuchi ◽  
Hirofumi Noguchi ◽  
Tohru Yagi ◽  
Shuji Hayashi
Keyword(s):  
Stem Cells ◽  
Adipogenic Differentiation ◽  
Three Dimensional ◽  
3D Culture ◽  
Human Stem Cells ◽  
Maintenance Medium ◽  
3D Cultures ◽  
Differentiation Medium ◽  
Human Ascs

The application of stem cells for cell therapy has been extensively studied in recent years. Among the various types of stem cells, human adipose tissue-derived stem cells (ASCs) can be obtained in large quantities with relatively few passages, and they possess a stable quality. ASCs can differentiate into a number of cell types, such as adipose cells and ectodermal cells. We therefore focused on the in vitro microenvironment required for such differentiation and attempted to induce the differentiation of human stem cells into microtissues using a microelectromechanical system. We first evaluated the adipogenic differentiation of human ASC spheroids in a three-dimensional (3D) culture. We then created the in vitro microenvironment using a 3D combinatorial TASCL device and attempted to induce the adipogenic differentiation of human ASCs. The differentiation of human ASC spheroids cultured in maintenance medium and those cultured in adipocyte differentiation medium was evaluated via Oil red O staining using lipid droplets based on the quantity of accumulated triglycerides. The differentiation was confirmed in both media, but the human ASCs in the 3D cultures contained higher amounts of triglycerides than those in the 2D cultures. In the short culture period, greater adipogenic differentiation was observed in the 3D cultures than in the 2D cultures. The 3D culture using the TASCL device with adipogenic differentiation medium promoted greater differentiation of human ASCs into adipogenic lineages than either a 2D culture or a culture using a maintenance medium. In summary, the TASCL device created a hospitable in vitro microenvironment and may therefore be a useful tool for the induction of differentiation in 3D culture. The resultant human ASC spheroids were “adipose-like microtissues” that formed spherical aggregation perfectly and are expected to be applicable in regenerative medicine as well as cell transplantation.

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