scholarly journals Tissue Harvesting Site Effect on the Canine Adipose Stromal Vascular Fraction Quantity and Quality

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 460
Author(s):  
Hanan Hendawy ◽  
Akiko Uemura ◽  
Danfu Ma ◽  
Ryosuke Namiki ◽  
Haney Samir ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Sampling Site ◽  
Stromal Vascular Fraction ◽  
Site Effect ◽  
Falciform Ligament ◽  
Great Promise ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Test ◽  
Clusters Of Differentiation

Mesenchymal stem cells (MSCs) constitute a great promise for regenerative therapy, but these cells are difficultly recovered in large amounts. A potent alternative is the stromal vascular fraction (SVF), non-cultured MSCs, separated from adipose tissue (AT). We aim to evaluate AT harvesting site effect on the SVF cells’ quantity and quality in dogs. Subcutaneous abdominal fat, falciform ligament and peri-ovarian fat were sampled. After SVF isolation, the trypan blue exclusion test and a hemocytometer were used to assess the cell viability and cellular yield. SVF cells were labeled for four surface antigenic markers, clusters of differentiation CD90, CD44, CD29, and CD45, and then examined by flow cytometry. Semi-quantitative RT-PCR was used to evaluate the gene expression of the former markers in addition to OCT-4 and CD34. SVF cells in the peri-ovarian AT recorded the highest viability% (99.63 ± 0.2%), as well as a significantly higher cellular yield (36.87 ± 19.6 × 106 viable cells/gm fat, p < 0.001) and a higher expression of adipose-derived mesenchymal stem cells AD-MSCs surface markers than that of other sites. SVF cells from the peri-ovarian site revealed a higher expression of MSC markers (CD90, CD44, and CD29) and OCT-4 compared to the other sites, with weak CD45 and CD34 expressions. The positive OCT-4 expression demonstrated the pluripotency of SVF cells isolated from different sites. To conclude, the harvesting site is a strong determinant of SVF cells’ quantity and quality, and the peri-ovarian site could be the best AT sampling site in dogs.

Culture and expansion of Adipose derived Mesenchymal stem cells in Ovine

2017 ◽  
Author(s):  
T. A. Kannan ◽  
J. Violet Beaulah ◽  
S. Ushakumary ◽  
B. Justin William ◽  
Geetha M. Ramesh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Trypan Blue ◽  
Povidone Iodine ◽  
Sterile Condition ◽  
Trypan Blue Exclusion ◽  
Tissue Samples ◽  
Trypan Blue Exclusion Test ◽  
Slaughter House

Culture and expansion of Adipose derived Mesenchymal stem cells (ADMSCs) in Ovine was carried out in this study. Ovine adipose tissue samples were collected from Chennai Corporation slaughter house under sterile condition in normal saline with five per cent povidone iodine, antibiotic and antimycotic solutions. Collected tissue samples were weighed and digested using collagenase enzyme to isolate adipose tissue derived mesenchymal stem cells (ADMSCs). Cell yield and viability of the cells were calculated by using trypan blue exclusion test. The cells were seeded at the density of one million cells in one T25 culture flask in Dulbecco’s modified Eagle’s medium-high glucose (DMEM-HG). On the day of seeding, the cells showed spherical morphology. Plastic adherence was noticed 24 hrs after seeding. Cell expansion was observed after 3 days. At P0 level, 70% confluency was attained on day 14 and the time taken to reach 70% confluency was reduced to 3-4 days in subsequent passages.

scholarly journals Regenerative Capacity of Adipose Derived Stem Cells (ADSCs), Comparison with Mesenchymal Stem Cells (MSCs)

2019 ◽  
Vol 20 (10) ◽  
pp. 2523 ◽  
Author(s):  
Loubna Mazini ◽  
Luc Rochette ◽  
Mohamed Amine ◽  
Gabriel Malka
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Therapy ◽  
Stromal Vascular Fraction ◽  
Large Cell ◽  
Great Promise ◽  
Adipose Derived Stem Cells ◽  
Clinical Practices ◽  
Cell Sources

Adipose tissue is now on the top one of stem cell sources regarding its accessibility, abundance, and less painful collection procedure when compared to other sources. The adipose derived stem cells (ADSCs) that it contains can be maintained and expanded in culture for long periods of time without losing their differentiation capacity, leading to large cell quantities being increasingly used in cell therapy purposes. Many reports showed that ADSCs-based cell therapy products demonstrated optimal efficacy and efficiency in some clinical indications for both autologous and allogeneic purposes, hence becoming considered as potential tools for replacing, repairing, and regenerating dead or damaged cells. In this review, we analyzed the therapeutic advancement of ADSCs in comparison to bone marrow (BM) and umbilical cord (UC)-mesenchymal stem cells (MSCs) and designed the specific requirements to their best clinical practices and safety. Our analysis was focused on the ADSCs, rather than the whole stromal vascular fraction (SVF) cell populations, to facilitate characterization that is related to their source of origins. Clinical outcomes improvement suggested that these cells hold great promise in stem cell-based therapies in neurodegenerative, cardiovascular, and auto-immunes diseases.

scholarly journals Derivation, characterization, and in vitro cell regeneration of canine white adipose tissue-derived mesenchymal stem cells obtained from a mesenteric region

2021 ◽  
Vol 82 (1) ◽  
Author(s):  
Anirban Mandal ◽  
Ajeet Kumar Jha ◽  
Dew Biswas ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
White Adipose Tissue ◽  
Stromal Vascular Fraction ◽  
Cell Regeneration ◽  
Wound Healing Assay ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background The study was conducted to assess the characterization, differentiation, and in vitro cell regeneration potential of canine mesenteric white adipose tissue-derived mesenchymal stem cells (AD-MSCs). The tissue was harvested through surgical incision and digested with collagenase to obtain a stromal vascular fraction. Mesenchymal stem cells isolated from the stromal vascular fraction were characterized through flow cytometry and reverse transcription-polymerase chain reaction. Assessment of cell viability, in vitro cell regeneration, and cell senescence were carried out through MTT assay, wound healing assay, and β-galactosidase assay, respectively. To ascertain the trilineage differentiation potential, MSCs were stained with alizarin red for osteocytes, alcian blue for chondrocytes, and oil o red for adipocytes. In addition, differentiated cells were characterized through a reverse transcription-polymerase chain reaction. Results We observed the elongated, spindle-shaped, and fibroblast-like appearance of cells after 72 h of initial culture. Flow cytometry results showed positive expression for CD44, CD90, and negative expression for CD45 surface markers. Population doubling time was found 18–24 h for up to the fourth passage and 30±0.5 h for the fifth passage. A wound-healing assay was used to determine cell migration rate which was found 136.9 ± 4.7 μm/h. We observed long-term in vitro cell proliferation resulted in MSC senescence. Furthermore, we also found that the isolated cells were capable of differentiating into osteogenic, chondrogenic, and adipogenic lineages. Conclusions Mesenteric white adipose tissue was found to be a potential source for isolation, characterization, and differentiation of MSCs. This study might be helpful for resolving the problems regarding the paucity of information concerning the basic biology of stem cells. The large-scale use of AD-MSCs might be a remedial measure in regenerative medicine.

scholarly journals Heterogeneity of mesenchymal stem cells in cartilage regeneration: from characterization to application

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Kangkang Zha ◽  
Xu Li ◽  
Zhen Yang ◽  
Guangzhao Tian ◽  
Zhiqiang Sun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Articular Cartilage ◽  
Cartilage Repair ◽  
Cartilage Damage ◽  
Cartilage Regeneration ◽  
Great Promise ◽  
Traditional Treatment ◽  
Different Types ◽  
Selection Of

AbstractArticular cartilage is susceptible to damage but hard to self-repair due to its avascular nature. Traditional treatment methods are not able to produce satisfactory effects. Mesenchymal stem cells (MSCs) have shown great promise in cartilage repair. However, the therapeutic effect of MSCs is often unstable partly due to their heterogeneity. Understanding the heterogeneity of MSCs and the potential of different types of MSCs for cartilage regeneration will facilitate the selection of superior MSCs for treating cartilage damage. This review provides an overview of the heterogeneity of MSCs at the donor, tissue source and cell immunophenotype levels, including their cytological properties, such as their ability for proliferation, chondrogenic differentiation and immunoregulation, as well as their current applications in cartilage regeneration. This information will improve the precision of MSC-based therapeutic strategies, thus maximizing the efficiency of articular cartilage repair.

scholarly journals DNA Methylation Changes duringIn VitroPropagation of Human Mesenchymal Stem Cells: Implications for Their Genomic Stability?

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Angela Bentivegna ◽  
Mariarosaria Miloso ◽  
Gabriele Riva ◽  
Dana Foudah ◽  
Valentina Butta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ex Vivo ◽  
Genomic Stability ◽  
Great Promise ◽  
Human Mscs ◽  
Low Oxygen ◽  
Long Term Culture ◽  
Functional Changes

Mesenchymal stem cells (MSCs) hold great promise for the treatment of numerous diseases. A major problem for MSC therapeutic use is represented by the very low amount of MSCs which can be isolated from different tissues; thusex vivoexpansion is indispensable. Long-term culture, however, is associated with extensive morphological and functional changes of MSCs. In addition, the concern that they may accumulate stochastic mutations which lead the risk of malignant transformation still remains. Overall, the genome of human MSCs (hMSCs) appears to be apparently stable throughout culture, though transient clonal aneuploidies have been detected. Particular attention should be given to the use of low-oxygen environment in order to increase the proliferative capacity of hMSCs, since data on the effect of hypoxic culture conditions on genomic stability are few and contradictory. Furthermore, specific and reproducible epigenetic changes were acquired by hMSCs duringex vivoexpansion, which may be connected and trigger all the biological changes observed. In this review we address current issues on long-term culture of hMSCs with a 360-degree view, starting from the genomic profiles and back, looking for an epigenetic interpretation of their genetic stability.

Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium

2021 ◽  
Author(s):  
Ajeet Kumar Jha ◽  
Anirban Mandal ◽  
Kalyani Ray ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stromal Vascular Fraction ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
High Plasticity ◽  
Rt Pcr ◽  
Differentiation Potential ◽  
Ovarian Surface ◽  
Isolation And Characterization

Background: Few studies have confirmed the presence of ovarian tissue stem cells indicating the capacity for differentiation. Based on this fact, it was hypothesized that mesenchymal stem cells (MSC) were found in ovarian surface epithelium (OSE) of canines that could easily be isolated. Methods: Both left and right ovaries were minced and digested using collagenase to obtain a stromal vascular fraction (SVF). MSCs were characterized using RT-PCR. To ascertain the trilineage differentiation potential, MSCs were stained with respective stain for osteocytes, chondrocytes and adipocytes. Result: We observed elongated, spindle-shaped and fibroblast like appearance of cells after 72 h of initial culture. Expression of MSC specific surface markers were observed through RT-PCR. Using Stem Pro® differentiation medium, OSE were differentiated into osteogenic, chondrogenic and adipogenic lineages and were found to be potential source for isolation, characterization and differentiation of MSCs. Canine (OSE) is easily accessible, multipotent and has high plasticity, holding promise for applications in regenerative medicine.

scholarly journals Adipose Derived-Mesenchymal Stem Cells Viability and Differentiating Features for Orthopaedic Reparative Applications: Banking of Adipose Tissue

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Ilaria Roato ◽  
Daniela Alotto ◽  
Dimas Carolina Belisario ◽  
Stefania Casarin ◽  
Mara Fumagalli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Articular Cartilage ◽  
Tissue Banking ◽  
Stromal Vascular Fraction ◽  
Single Infusion ◽  
Knee Arthritis ◽  
Symptomatic Knee

Osteoarthritis is characterized by loss of articular cartilage also due to reduced chondrogenic activity of mesenchymal stem cells (MSCs) from patients. Adipose tissue is an attractive source of MSCs (ATD-MSCs), representing an effective tool for reparative medicine, particularly for treatment of osteoarthritis, due to their chondrogenic and osteogenic differentiation capability. The treatment of symptomatic knee arthritis with ATD-MSCs proved effective with a single infusion, but multiple infusions could be also more efficacious. Here we studied some crucial aspects of adipose tissue banking procedures, evaluating ATD-MSCs viability, and differentiation capability after cryopreservation, to guarantee the quality of the tissue for multiple infusions. We reported that the presence of local anesthetic during lipoaspiration negatively affects cell viability of cryopreserved adipose tissue and cell growth of ATD-MSCs in culture. We observed that DMSO guarantees a faster growth of ATD-MSCs in culture than trehalose. At last, ATD-MSCs derived from fresh and cryopreserved samples at −80°C and −196°C showed viability and differentiation ability comparable to fresh samples. These data indicate that cryopreservation of adipose tissue at −80°C and −196°C is equivalent and preserves the content of ATD-MSCs in Stromal Vascular Fraction (SVF), guaranteeing the differentiation ability of ATD-MSCs.

scholarly journals BMAC and Adipose-Derived MSCs Treatment for Knee Osteoarthritis: A Systematic Review

2021 ◽  
Vol 7 (04) ◽  
pp. 01-11
Author(s):  
Pooja Pithadia
Keyword(s):  
Systematic Review ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Knee Osteoarthritis ◽  
Bone Marrow Aspirate ◽  
Stromal Vascular Fraction ◽  
Cochrane Library ◽  
Bone Marrow Aspirate Concentrate ◽  
Mesh Terms

Background: Knee osteoarthritis is the most common musculoskeletal progressive disorder that affects nearly 303 million people worldwide. This condition prevails in 10% males and 13% females among the elders above 60. Although there is conventional nonsurgical and surgical treatment available for knee osteoarthritis, there is a fascinating interest in bone marrow aspirate concentrate (BMAC) as well as adipose-derived mesenchymal stem cells (AD-MSC), including enzymatically treated stromal vascular fraction (SVF) and mechanically treated (microfat/nanofat) injections among physicians. Hence, this systematic review aims to determine the efficacy of BMAC and AD-MSCs (enzyme and mechanically treated) injections for knee osteoarthritis treatment. Methods: A systematic review was performed on the following data sources (PubMed, Scopus, Google Scholar, EMBASE, and Cochrane Library) published on March 31, 2021. The keywords or MeSH terms include 'Knee Osteoarthritis with 'Bone marrow aspirate concentrate' OR 'BMAC' or with 'Adipose-derived mesenchymal stem cells (AD-MSC)' or with 'Stromal vascular fraction' OR 'SVF' or 'Mechanically treated AD-MSC (mfat/nanofat)'. In addition, the retrieved articles were further reviewed to identify relevant research studies. Results: The authors reviewed and tabulated data based on the year of study, study type, therapy protocol, patient population, outcome measures, and interpretation. Among the 382 records screened, 43 studies (16 on BMAC and 27 on AD-MSCs) were included in the systematic review study. Among them, only 5 were randomized controlled trials. These selected studies demonstrated short-term positive outcomes such as improvement in knee pain and function with no adverse side effects. Moreover, researchers reported varied administration methods of BMAC or AD-MSC either as standalone or in combination with other conservative procedures such as PRP (Platelets Rich Plasma), HA (Hyaluronic acid), or surgery. Conclusions: BMAC and AD-MSC (enzymatically and mechanically treated) injections prove safer and more efficacious in patients with knee osteoarthritis for a shorter duration of 2 years. However, the available literature lacks high-quality studies with no varied clinical settings and long-term follow-up of more than two years.

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