Effect of Mineral Pitch on the Proliferation of Human Adipose Derived Stem Cells on Acellular Scaffold

Purpose : Acellular scaffold extracted from extracellular matrix (ECM) have been used for constructive and regenerative medicine. Adipose derived stem cells (ADSCs) can enhance the vascularization capacity of scaffolds. High mobility group box 1 (HMGB1) and stromal derived factor1 (SDF1) are considered as two important factors in vascularization and immunologic system. In this study, the effect of mineral pitch on the proliferation of human ADSCs was evaluated. In addition to HMGB1 and SDF1, factors expression in acellular scaffold was also assessed. Methods: To determine acellular scaffold morphology and the degree of decellularization, hematoxylin & eosin (H&E), 6-diamidino-2-phenylindole (DAPI), and Masson’s trichrome staining were applied. The scaffolds were treated with mineral pitch. Also, ADSCs were seeded on the scaffolds, and adhesion of the cells to the scaffolds were assessed using field emission scanning electron microscopy (FE-SEM). In addition, the efficiency of mineral pitch to induce the proliferation of ADSCs on the scaffolds was evaluated using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. To measure HMGB1 and SDF1 mRNA expression, real-time polymerase chain reactions (RT-PCR) was used. Results: FE-SEM showed that decellularized matrix possesses similar matrix morphology with a randomly oriented fibrillar structure and interconnecting pores. No toxicity was observed in all treatments, and cell proliferation were supported in scaffolds. The important point is that, the proliferation capacity of ADSCs on Mineral pitch loaded scaffolds significantly increased after 48 h incubation time compared to the unloaded scaffold (P<0.001). Conclusion: The results of this study suggest that mineral pitch has potentials to accelerate proliferation of ADSCs on the acellular scaffolds.

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The angiogenic properties of human adipose-derived stem cells (HASCs) are modulated by the High mobility group box protein 1 (HMGB1)

International Journal of Cardiology ◽

10.1016/j.ijcard.2017.09.165 ◽

2017 ◽

Vol 249 ◽

pp. 349-356 ◽

Cited By ~ 4

Author(s):

Federico Biscetti ◽

Stefano Gentileschi ◽

Flavio Bertucci ◽

Maria Servillo ◽

Vincenzo Arena ◽

...

Keyword(s):

Stem Cells ◽

High Mobility ◽

High Mobility Group ◽

Adipose Derived Stem Cells

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Influence of Age and Body Mass Index on the Yield and Proliferation Capacity of Adipose-Derived Stem Cells

Aesthetic Plastic Surgery ◽

10.1007/s00266-011-9743-7 ◽

2011 ◽

Vol 35 (6) ◽

pp. 1097-1105 ◽

Cited By ~ 28

Author(s):

Ali Mojallal ◽

Charlotte Lequeux ◽

Christo Shipkov ◽

Antoine Duclos ◽

Fabienne Braye ◽

...

Keyword(s):

Body Mass Index ◽

Stem Cells ◽

Body Mass ◽

Adipose Derived Stem Cells ◽

Proliferation Capacity

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COMPARISON OF PROLIFERATION AND IMMUNOPHENOTYPE OF MSC, OBTAINEDFROM BONE MARROW, ADIPOSE TISSUE AND UMBILICAL CORD

HERALD of North-Western State Medical University named after I I Mechnikov ◽

10.17816/mechnikov20157214-22 ◽

2015 ◽

Vol 7 (2) ◽

pp. 14-22

Author(s):

A A Aisenstadt ◽

N I Enukashvili ◽

T L Zolina ◽

L V Alexandrov ◽

A B Smoljaninov

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Adipose Tissue ◽

Clinical Practice ◽

Umbilical Cord ◽

Adipose Derived Stem Cells ◽

Non Invasive ◽

Bone Marrow Adipose Tissue ◽

Proliferation Capacity ◽

Marrow Adipose Tissue

Mesenchymal stem cells (MSC) can be applied for treatment of different diseases. Human MSC have been isolated from bone marrow, adipose tissue, umbilical cord blood. umbilical cord MSCs (uC- MSC) are obtained during birth with non-invasive, non-traumatic methods and thus seem a good candidate for clinical practice instead of bone-marrow MSC (BM-MSC). It is yet unknown wheth- er the immunophenotype and proliferation capacity of uC-MSC are similar to adipose-derived stem cells (AdSCs) or BM-MSC. The goal of this research was to study the immunophenotype and prolifer- atiion capacity of uC-MSC, AdSCs and BM-MSC. The results indicated that MSC of different origin had similar morphological and immunophenotypic characteristics with minor differences. uC-MSC differed from other cultures by constant level of Cd105 expression, the presence of minor Cd10 - and Cd13 - populations and higher proliferative activity. BM-MSC were characterized by reduced expres- sion levels of Cd90, compared with the uC-MSCs and AdSCs. These data confirm the similarity of uC-MSC with BM-MSCs and AdSCs and the possibility of their use in clinical practice instead of hard-to-obtain BM-MSC.

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O49: EFFECT OF AFIMOXIFENE (Z-4-HYDROXYTAMOXIFEN) ON ADIPOSE-DERIVED STEM CELLS

British Journal of Surgery ◽

10.1093/bjs/znab117.049 ◽

2021 ◽

Vol 108 (Supplement_1) ◽

Author(s):

RS Challapalli ◽

RM Dwyer ◽

N McInerney ◽

MJ Kerin ◽

AJ Lowery

Keyword(s):

Breast Cancer ◽

Stem Cells ◽

Adipose Tissue ◽

Breast Reconstruction ◽

Cancer Patients ◽

Hormonal Therapy ◽

Deleterious Effect ◽

Adipose Derived Stem Cells ◽

Breast Cancer Patients ◽

Human Adscs

Abstract Background Adipose-derived stem cells (ADSCs) are a promising cell source for adipose tissue engineering. Currently, novel breast reconstruction techniques using ADSCs are actively being explored. Systemic chemo- & hormonal therapy may impede tissue regeneration in breast cancer patients. However, the effect of hormonal therapy on ADSCs and their regenerative capabilities is poorly understood. Aims The current study aims to analyse the effect of Afimoxifene, an active metabolite of Tamoxifen, on proliferation and viability of ADSCs in vitro Method Lipoaspirates or adipose tissue were obtained with informed consent from breast cancer patients and healthy controls. The isolated ADSCs were subjected to single or multiple dose(s) of Z-4-Hydroxytamoxifen (12.5nM, 25nM, 50nM, 100nM and 1μM) and, analyzed on days 1, 3 and 5, using CellTitre 96 ® AQueous Cell proliferation assay. T47D and MDA-MB-231 cell lines were used as positive and negative controls, respectively. Result ADSCs were obtained from a total of 3 patients. ADSCs were isolated from a cancer patient with/without active disease at the time of sample procurement and healthy subjects opting for elective cosmetic procedures. Morphology, CFU assay and adipogenic differentiation assay were used to validate stem cell population. Afimoxifene demonstrated no statistically significant reduction in the viability or proliferation of the ADSCs, irrespective of the dose- or time-dependent exposure or cancer status (p >0.05). Conclusion Afimoxifene has no deleterious effect on viability or proliferation of human ADSCs up to 1μM concentration. Key words Adipose-derived stem cells, Afimoxifene, Breast reconstruction, Hormone therapy, Tamoxifen. Take-home message Afimoxifene has no deleterious effect on viability or proliferation of human ADSCs up to 1μM concentration

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Human adipose-derived stem cells combined with high mobility group box protein 1 might be a novel therapeutic strategy for the treatment of peripheral arterial disease

International Journal of Cardiology ◽

10.1016/j.ijcard.2018.06.085 ◽

2019 ◽

Vol 274 ◽

pp. 304

Author(s):

Changhui Zhou ◽

Ying Mu ◽

Weihua Wang

Keyword(s):

Stem Cells ◽

Peripheral Arterial Disease ◽

Therapeutic Strategy ◽

High Mobility ◽

Arterial Disease ◽

High Mobility Group ◽

Adipose Derived Stem Cells ◽

Peripheral Arterial ◽

Novel Therapeutic

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The combination of trehalose and glycerol: an effective and non-toxic recipe for cryopreservation of human adipose-derived stem cells

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01969-0 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Tian-Yu Zhang ◽

Poh-Ching Tan ◽

Yun Xie ◽

Xiao-Jie Zhang ◽

Pei-Qi Zhang ◽

...

Keyword(s):

Stem Cells ◽

Cell Viability ◽

High Efficiency ◽

Cell Function ◽

Clinical Applications ◽

Adipose Derived Stem Cells ◽

Cryoprotective Agents ◽

Proliferation Capacity ◽

Cell Based Therapy ◽

Toxic Concentrations

Abstract Background Adipose-derived stem cells (ADSCs) promote tissue regeneration and repair. Cryoprotective agents (CPAs) protect cells from cryodamage during cryopreservation. Safe and efficient cryopreservation of ADSCs is critical for cell-based therapy in clinical applications. However, most CPAs are used at toxic concentrations, limiting their clinical application. Objective The aim of this study is to develop a non-toxic xeno-free novel CPA aiming at achieving high-efficiency and low-risk ADSC cryopreservation. Methods We explored different concentrations of trehalose (0.3 M, 0.6 M, 1.0 M, and 1.25 M) and glycerol (10%, 20%, and 30% v/v) for optimization and evaluated and compared the outcomes of ADSCs cryopreservation between a combination of trehalose and glycerol and the commonly used CPA DMSO (10%) + FBS (90%). All samples were slowly frozen and stored in liquid nitrogen for 30 days. The effectiveness was evaluated by the viability, proliferation, migration, and multi-potential differentiation of the ADSCs after thawing. Results Compared with the groups treated with individual reagents, the 1.0 M trehalose (Tre) + 20% glycerol (Gly) group showed significantly higher efficiency in preserving ADSC activities after thawing, with better outcomes in both cell viability and proliferation capacity. Compared with the 10% DMSO + 90% FBS treatment, the ADSCs preserved in 1.0 M Tre + 20% Gly showed similar cell viability, surface markers, and multi-potential differentiation but a significantly higher migration capability. The results indicated that cell function preservation can be improved by 1.0 M Tre + 20% Gly. Conclusions The 1.0 M Tre + 20% Gly treatment preserved ADSCs with a higher migration capability than 10% DMSO + 90% FBS and with viability higher than that with trehalose or glycerol alone but similar to that with 10% DMSO + 90% FBS and fresh cells. Moreover, the new CPA achieves stemness and multi-potential differentiation similar to those in fresh cells. Our results demonstrate that 1.0 M Tre + 20% Gly can more efficiently cryopreserve ADSCs and is a non-toxic CPA that may be suitable for clinical applications.

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The Identification of HSA-MIR-17-5P Existence in the Exosome of Adipose-Derived Stem Cells and Adipocytes

Journal of Biomimetics Biomaterials and Biomedical Engineering ◽

10.4028/www.scientific.net/jbbbe.52.66 ◽

2021 ◽

Vol 52 ◽

pp. 66-75

Author(s):

Sinta Murlistyarini ◽

Lulus Putri Aninda ◽

Ufida Aini Afridafaz ◽

Sri Widyarti ◽

Agustina Tri Endharti ◽

...

Keyword(s):

Stem Cells ◽

Pregnant Woman ◽

Rna Isolation ◽

Cell Types ◽

Adipose Derived Stem Cells ◽

Gene Expressions ◽

Human Adscs ◽

Elective Cesarean ◽

Questionable Existence

MicroRNAs (miRNAs) have ability to down-regulate gene expressions. hsa-miR-17-5p, has been confirmed as an oncogene or tumor suppressor. However, the existence on human adipose-derived stem cells (ADSCs) or adipocytes, is still unclear. Many researchers emphasizing the role of hsa-miR-17-5p on cellular senescence, aging and cancer, but not specific on the expression in the exosome of human ADSCs and adipocytes. The primary ADSCs were derived from subcutaneous adipose tissue of pregnant woman during elective cesarean operation, then processed by combining conventional and enzymatic methods. Adipocytes were differentiated by using the StemPro Adipogenesis Differentiation kit® and Oil Red-O staining. Exosomes were isolated using Exosome Purification and RNA Isolation kit® and were characterized by scanning electron microscope. The markers, CD34 and CD44, were identified and analyzed by using FACS analysis. Subsequently, microRNA was extracted and observed for hsa-miR-17-5p expression. This study showed that ADSCs and adipocytes were proved to express CD34+ and CD44+. The hsa-miR-17-5p were also detected in both the exosome of ADSCs and adipocytes. Although the source of the ADSCs was from pregnant woman, the characteristic was similar with the ones from non-pregnant woman. Our study also supports the questionable existence of CD34 in ADSCs. Having confirmed the characteristics, we proved that the exosomes of ADSCs and adipocytes expressed similar hsa-miR-17-5p despite they are from phenotypically different cell types and may have distinct roles. However, further research steps should be done in the future to verify the role of hsa-miR-17-5p towards senescent cell and ADSC differentiation.

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The Combination of Trehalose and Glycerol: An Effective and Non-Toxic Recipe for Cryopreservation of Human Adipose-Derived Stem Cells

10.21203/rs.3.rs-60477/v2 ◽

2020 ◽

Author(s):

Tian-Yu Zhang ◽

Poh-Ching Tan ◽

Yun Xie ◽

Xiao-Jie Zhang ◽

Pei-Qi Zhang ◽

...

Keyword(s):

Stem Cells ◽

Cell Viability ◽

High Efficiency ◽

Cell Function ◽

Clinical Applications ◽

Adipose Derived Stem Cells ◽

Cryoprotective Agents ◽

Proliferation Capacity ◽

Cell Based Therapy ◽

Toxic Concentrations

Abstract Background: Adipose-derived stem cells (ADSCs) promote tissue regeneration and repair. Cryoprotective agents (CPAs) protect cells from cryodamage during cryopreservation. Safe and efficient cryopreservation of ADSCs is critical for cell-based therapy in clinical applications. However, most CPAs are used at toxic concentrations, limiting their clinical application. Objective: The aim of this study is to develop a non-toxic xeno-free novel CPA aiming at achieving high-efficiency and low-risk ADSC cryopreservation.Methods: We explored different concentrations of trehalose (0.3 M, 0.6 M, 1.0 M, and 1.25 M) and glycerol (10%, 20%, and 30% v/v) for optimization and evaluated and compared the outcomes of ADSCs cryopreservation between a combination of trehalose and glycerol and the commonly used CPA DMSO (10%) + FBS (90%). All samples were slowly frozen and stored in liquid nitrogen for 30 days. The effectiveness was evaluated by the viability, proliferation, migration and multi-potential differentiation of the ADSCs after thawing. Results: Compared with the groups treated with individual reagents, the 1.0 M Tre + 20% Gly group showed significantly higher efficiency in preserving ADSC activities after thawing, with better outcomes in both cell viability and proliferation capacity. Compared with the 10% DMSO + 90% FBS treatment, the ADSCs preserved in 1.0 M Tre + 20% Gly showed similar cell viability, surface markers and multi-potential differentiation but a significantly higher migration capability. The results indicated that cell function preservation can be improved by 1.0 M Tre + 20% Gly. Conclusions: The 1.0 M Tre + 20% Gly treatment preserved ADSCs with a higher migration capability than 10% DMSO + 90% FBS and with viability higher than that with trehalose or glycerol alone but similar to that with 10% DMSO + 90% FBS and fresh cells. Moreover, the new CPA achieves stemness and multi-potential differentiation similar to those in fresh cells. Our results demonstrate that 1.0 M Tre + 20% Gly can more efficiently cryopreserve ADSCs and is a non-toxic CPA that may be suitable for clinical applications.

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Generation of Neurospheres from Human Adipose-Derived Stem Cells

BioMed Research International ◽

10.1155/2015/743714 ◽

2015 ◽

Vol 2015 ◽

pp. 1-10 ◽

Cited By ~ 11

Author(s):

Erfang Yang ◽

Na Liu ◽

Yingxin Tang ◽

Yang Hu ◽

Ping Zhang ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Adult Brain ◽

Adipose Derived Stem Cells ◽

Alternative Source ◽

Multipotent Stem Cells ◽

Human Adscs ◽

Epidermal Growth ◽

Polymerase Chain ◽

Proliferative Ability

Transplantation of neural stem cells (NSCs) to treat neurodegenerative disease shows promise; however, the clinical application of NSCs is limited by the invasive procurement and ethical concerns. Adipose-derived stem cells (ADSCs) are a source of multipotent stem cells that can self-renew and differentiate into various kinds of cells; this study intends to generate neurospheres from human ADSCs by culturing ADSCs on uncoated culture flasks in serum-free neurobasal medium supplemented with B27, basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF); the ADSCs-derived neurospheres were terminally differentiated after growth factor withdrawal. Expression of Nestin, NeuN, MAP2, and GFAP in ADSCs and terminally differentiated neurospheres was shown by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting, and immunocytochemistry; cell proliferation in neurospheres was evaluated by cell cycle analyses, immunostaining, and flow cytometry. These data strongly support the conclusion that human ADSCs can successfully differentiate into neurospheres efficiently on uncoated culture flasks, which present similar molecular marker pattern and proliferative ability with NSCs derived from embryonic and adult brain tissues. Therefore, human ADSCs may be an ideal alternative source of stem cells for the treatment of neurodegenerative diseases.

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