Adipose-derived stem cells in the treatment of knee osteoarthritis: from extraction methods to the preparation of the transplant

Introduction: Recent studies have investigated the use of adipose tissue as source of mesenchymal stem cells in the treatment of knee osteoarthritis in humans. However, there are still several protocols being performed. Objective: Analyze the protocols published in the literature in the last ten years and to investigate how they are being carried out and if they are following the criteria adopted by the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Methodology: Articles from the PubMed, ScienceDirect and Lilacs database published in January / 2010 until the present time, which were evaluated in order to investigate the use of adipose-derived stem cells in the treatment of knee osteoarthritis. Results: Thirty four articles were evaluated in its entiraty. The abdominal area was the most choosen to do the liposuction, however the quantities of adipose tissue removed and the number of cells transplanted was variable. It is hightlited the enzimatic digestion of adipose tissue with collagenase as extraction method. Only 14 articles complied all the 3 criteria required to prove the real presence of mesenchymal stem cells in the samples that was transplanted. However, all the articles showed improvement of function and pain. Final considerations: Thus, even the results found are promising, the evidence is still limited in humans and the variability of the methodology makes it difficult to standardize the technique, also its implementation as a reference in the treatment of knee osteoarthritis.

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Are Adipose-Derived Stem Cells from Liver Falciform Ligaments Another Possible Source of Mesenchymal Stem Cells?

Cell Transplantation ◽

10.3727/096368916x693833 ◽

2017 ◽

Vol 26 (5) ◽

pp. 855-866 ◽

Cited By ~ 4

Author(s):

Sang Woo Lee ◽

Jae Uk Chong ◽

Seon Ok Min ◽

Seon Young Bak ◽

Kyung Sik Kim

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Liver Disease ◽

Subcutaneous Adipose Tissue ◽

Falciform Ligament ◽

Pcr Analysis ◽

Marker Genes ◽

Adipose Derived Stem Cells ◽

Subcutaneous Adipose

Falciform ligaments in the liver are surrounded by adipose tissue. We investigated the capability of adipose-derived stem cells from human liver falciform ligaments (hLF-ADSCs) to differentiate into hepatic-type cells and confirmed the functional capacity of the cells. Mesenchymal stem cells (MSCs) were isolated from the liver falciform ligament and abdominal subcutaneous adipose tissue in patients undergoing partial hepatectomy for liver disease. Cells were cultivated in MSC culture medium. Properties of MSCs were confirmed by flow cytometry, RT-PCR analysis, immunocytochemistry assays, and multilineage differentiation. Hepatic induction was performed using a three-step differentiation protocol with various growth factors. Morphology, capacity for expansion, and characteristics were similar between hLF-ADSCs and adipose-derived stem cells from human abdominal subcutaneous adipose tissue (hAS-ADSCs). However, hematopoietic– and mesenchymal–epithelial transition (MET)-related surface markers (CD133, CD34, CD45, and E-cadherin) had a higher expression in hLF-ADSCs. The hepatic induction marker genes had a higher expression in hLF-ADSCs on days 7 and 10 after the hepatic induction. Albumin secretion was similar between hLF-ADSCs and hAS-ADSCs at 20 days after the hepatic induction. The hLF-ADSCs had a different pattern of surface marker expression relative to hAS-ADSCs. However, proliferation, multilineage capacity, and hepatic induction were similar between the cell types. Accordingly, it may be a useful source of MSCs for patients with liver disease.

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Comparison between Mesenchymal Stem Cells obtained from Bone Marrow, Adipose Tissue and Wharton Gelatin according to the ISCT Criteria

10.17488/rmib.30.1.23 ◽

2017 ◽

Author(s):

◽

A. Parra-Barrera

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Cellular Therapy ◽

Wharton’S Jelly ◽

Alternative Source ◽

Wharton's Jelly ◽

Bone Marrow Adipose Tissue ◽

Marrow Adipose Tissue

Mesenchymal stem cells (MSC) represent a heterogeneous population with the capacity to self-renew and differentiate into different cell types. At the middle of the last century these cells initially were described in bone marrow (BM), thence this tissue has become the gold standard for obtaining and characterization of MSC. It is known that these cells are housed in specific areas called niches distributed throughout all body, where they contribute to tissue regeneration processes of self-tissue were they are located. However, finding an alternative source of CTM with the same characteristics that have showed in MO, but its obtention no represent a risk since the donor is essential to their use for therapeutic purposes. In this study we isolated mesenchymal stem cells from bone marrow, adipose tissue and Wharton’s jelly and they were compared in their characteristics in according to the standards of the International Society for Cellular Therapy (ISCT). The results showed that the morphology as well as adipogenic and osteogenic differentiation and also the expression of surface antigens (CD90, CD73, and CD105) from all tissues accomplished the standards, although Wharton’s jelly represented the best option.

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Comparison between Mesenchymal Stem Cells obtained from Bone Marrow, Adipose Tissue and Wharton Gelatin according to the ISCT Criteria

10.17488/rmib.381.23 ◽

2017 ◽

Author(s):

◽

A. Parra-Barrera ◽

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Cellular Therapy ◽

Wharton’S Jelly ◽

Alternative Source ◽

Wharton's Jelly ◽

Bone Marrow Adipose Tissue ◽

Marrow Adipose Tissue

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Use of microfiltered vs only disaggregated mesenchymal stem cells from adipose tissue in regenerative medicine

SCRIPTA MEDICA ◽

10.5937/scriptamed51-24968 ◽

2020 ◽

Vol 51 (3) ◽

pp. 152-157

Author(s):

Fabiano Svolacchia ◽

Lorenzo Svolacchia

Keyword(s):

Stem Cells ◽

Flow Cytometry ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Bioactive Peptides ◽

Clinical Use ◽

Adult Mesenchymal Stem Cells ◽

Epidermal Regeneration ◽

Inflammatory Component ◽

Number Of Cells

Background: Clinical use of adult mesenchymal stem cells (MSCa) in medicine and regenerative surgery is constantly evolving. Adipose tissue-derived stem cells (ADSc) are capable of inducing the production of new extracellular matrix (ECM), deposition of new collagen and early revascularisation. Methods: Flow cytometry was performed for 2 mL of cell colonies harvested from adipose tissue (AT). Comparation has been made of at disaggregated only and the same at disaggregated and microfiltered at 50 mm, 100 mm and 200 mm. Signs of inflammation after dermo-epidermal regeneration session through the mesotherapy method were observed and compared. Results: Even after filtration, significant number of ADSc was collected. An increase in the size of the filter did not always translate into an increase in the number of cells that were found in the microfiltrate. In the non-filtered at disaggregated in both cases, highest number of cells was found, as expected, but at the expense of more pronounced inflammation. Sampling with the 16 Gauge needle produces superior results compared to the cannula in all cases. Conclusion: With this method in medicine and regenerative surgery it will be easier to exploit the growth factors, mRNA, MicroRNA, lipids and bioactive peptides emitted in the MSCa signalling micro-vesicles as they are isolated from the inflammatory component.

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Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT)

Cytotherapy ◽

10.1016/j.jcyt.2013.02.006 ◽

2013 ◽

Vol 15 (6) ◽

pp. 641-648 ◽

Cited By ~ 897

Author(s):

Philippe Bourin ◽

Bruce A. Bunnell ◽

Louis Casteilla ◽

Massimo Dominici ◽

Adam J. Katz ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

International Society ◽

Stromal Cells ◽

Cellular Therapy ◽

Stromal Vascular Fraction ◽

International Federation ◽

Joint Statement ◽

Stromal Stem Cells

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Treatment of Oral Ulcers in Dogs Using Adipose Tissue-Derived Mesenchymal Stem Cells

Journal of Clinical Pediatric Dentistry ◽

10.17796/jcpd.38.3.193115427jg6vl60 ◽

2014 ◽

Vol 38 (3) ◽

pp. 215-222 ◽

Cited By ~ 4

Author(s):

N M Alamoudi ◽

E A El Ashiry ◽

N M Farsi ◽

D A El Derwi ◽

H M Atta

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Growth Factor ◽

Ulcer Healing ◽

Cellular Therapy ◽

Healing Process ◽

Oral Ulcer ◽

Oral Ulcers

Aim: Adipose tissue Derived Mesenchymal Stem cells (ADMSCs) represent a promising tool for new clinical concepts in supporting cellular therapy. The goal of this study was to investigate the effects of ADMSCs transplantation on oral ulcer healing in dogs. Study design: Mesenchymal stem cells were isolated from adipose tissues of dogs obtained by suction-assisted lipectomy (liposuction), by dish adherence and were expanded in culture. Oral ulcers were induced by topical application of formocresol in the oral cavity of 18 dogs. The dogs were classified into 3 groups. Either autologous ADMSCs, Corticosteriod (Dexamethasone) or vehicle (saline) was injected. The healing process of the ulcer was monitored histopathologically. Gene expression of vascular endothelial growth factor (VEGF), platelets derived growth factor (PDGF), epidermal growth factor (EGF) and collagen was assessed in biopsies obtained from all ulcers ‘’as healing markers’’, by real time polymerase chain reaction (PCR). Results: ADMSCs group showed significantly accelerated oral ulcer healing compared with the Dexamethasone and control groups. There was increased expression of VEGF, PDGF, EGF and collagen genes in ADMSCs-treated ulcers compared with Dexamethasone and controls. Conclusion: ADMSCs transplantation may help accelerate oral ulcer healing, possibly through the induction of angiogenesis by VEGF and PDGF, as well as epithelial and connective tissue proliferation as evidenced by increased EGF and collagen gene expression.

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Expression of CD34 marker in the adipose tissue derived stem cells

Progress in Stem Cell ◽

10.15419/psc.v5i1.399 ◽

2018 ◽

Vol 5 (1) ◽

Author(s):

Phuc Van Pham ◽

Ngoc Bich Vu ◽

Van Hong Tran

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Flow Cytometry ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Hematopoietic Stem Cells ◽

Adipose Derived Stem Cells ◽

Differentiation Potential ◽

Hematopoietic Stem ◽

Cd34 Expression

Introduction: Adipose-derived stem cells (ADSCs) are considered as mesenchymal stem cells (MSCs). Indeed, they display all characteristics of MSCs that compliant with the minimal criteria of MSCs suggested by Domonici et al. (2006). However, some recent studies showed that ADSCs contain the subpopulation that was positive with CD34 marker – a marker of hematopoietic stem cells. This study aimed to analyze and determine the expression of CD34 marker in ten samples of ADSCs obtained from 10 donors. Methods: All ADSC samples were isolated and expanded according to the published previous protocols. They were confirmed as the MSCs with some markers and differentiation potential, excepting the CD34 expression. Then they were cultured and analyzed the expression of CD34 by flow cytometry at passage 3, 5, 7 and 9. Results: The results showed that expression of CD34 in ADSCs was different between donors and their passages that accounted from 1.21% to 23.38%. Conclusion: These results suggested that ADSCs are not ‘truly” MSCs like MSCs from bone marrow.

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Comparative Study of Expansion and Proliferation of Adult Mice Mesenchymal Stem Cells Derived from Bone Marrow and Adipose Tissue

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2016.10.2.481 ◽

2016 ◽

Vol 10 (2) ◽

pp. 39-46

Author(s):

Mahfoodha Abbas Umran ◽

Marwa Ibrahim Salman

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Success Rate ◽

Cellular Therapy ◽

Immune Phenotype ◽

Adult Mice ◽

Promising Source ◽

Proliferation Period

The importance of Mesenchymal stem cells (MSCs) represents a favorable tool for new clinical concepts in supporting tissue engineering and cellular therapy. Bone marrow (BM) was considered important source contain mesenchymal stem cells .Another promising source of MSCs is adipose tissue (AT). MSCs derived from these sources compared regarding morphology, the success rate of isolating MSCs, expansion potential by rate of colony forming and immune phenotype. The obtained results from this study showed no obvious considerable differences concerning the morphology and immune phenotype of the MSCs derived from these sources were obvious. Differences observed concerning to the success rate of isolating MSCs, which was approximately more than 90% for BM, while it reached about 70% for AT after seven days of culturing, as well as the rate of colony forming was lower in AT cells in comparison to that obtained in BM at the same period. However, AT-MSCs could be required longest time to complete monolayer confluence, whereas BM-MSCs had the shortest proliferation period. Cells from both sources determined according to immunohistochemistry by CD105+ and CD34.¯ Conclusions revealed that MSCs can easily and successfully obtained from bone marrow and adipose tissues, and both tissues appears suitable sources of stem cells for potential use in regenerative medicine, repairing damaged tissue nevertheless the BM-MSCs more effectual in expansion and proliferation.

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GMP-Compliant Human Adipose Tissue-Derived Mesenchymal Stem Cells for Cellular Therapy

Methods in Molecular Biology - Stem Cells and Good Manufacturing Practices ◽

10.1007/7651_2014_112 ◽

2014 ◽

pp. 93-107 ◽

Cited By ~ 9

Author(s):

Hamid-Reza Aghayan ◽

Parisa Goodarzi ◽

Babak Arjmand

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Cellular Therapy ◽

Human Adipose Tissue

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308 ISOLATION, DIFFERENTIATION, AND IMMUNOPHENOTYPIC CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM EQUINE ADIPOSE TISSUE AND BONE MARROW

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab308 ◽

2011 ◽

Vol 23 (1) ◽

pp. 250

Author(s):

E. Iacono ◽

B. Merlo ◽

A. Spadari ◽

G. Mari ◽

F. Ricci ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Cellular Therapy ◽

Population Doubling ◽

Blue Staining ◽

Alcian Blue Staining ◽

Alizarin Red ◽

Minimum Criteria

Minimum criteria for the characterisation of human mesenchymal stem cells (MSC) are a) adhesion to the plastic when maintained under culture conditions; b) expression of CD105, CD73, and CD90, and no expression for CD45, CD34, and CD14; and c) differentiation into osteoblasts, adipocytes, and chondroblasts in vitro. One major difficulty in characterising equine MSC is the absence of specific monoclonal antibodies and evidence that certain markers from other species do not cross-react with the equine species. The aim of this work was to isolate, cultivate, differentiate, and conduct cellular characterisation of MSC derived from equine adipose tissue (AT) and bone marrow (BM). Adipose tissue collection was performed at the base of the horses’ tails, and BM was aspirated from the iliac crest. Mononuclear cell fraction was isolated and cultured as previously described by (Colleoni et al. 2009 Vet. Res. Commun. 33, 811–821). Chondrogenic, osteogenic, and adipogenic differentiation were performed in monolayer culture, and evidence for differentiation was made by morphological and cytological evaluations. For molecular characterisation, cells were treated with trypsin, washed with PBS, and fixed with Reagent 1 (Intraprep Kit, Beckman Coulter, Miami, FL, USA), following the manufacturer’s instructions. Samples, after washings, were incubated for 20 min at room temperature with CD105, CD90, CD44, CD45, CD34, CD14, and CD73 mAbs, directly conjugated to fluorescein isothiocyanate, PE, or APC (Beckman Coulter). Appropriate conjugate isotype controls were applied (Beckman Coulter). After staining, cells were washed twice with PBS, and fluorescence intensity was evaluated with a FC500 two-laser equipped cytometer (Beckman Coulter). Results were further analysed with the CXP dedicated program. Samples volumes were 68 ± 23.6 mL for BM and 5.6 ± 1.1 g for AT; in both AT and BM, the isolation rate was 100% (AT: 4/4; BM: 5/5). Undifferentiated cells were passaged up to 8 times for AT and 5 times for BM; population-doubling times (DT) were calculated, and data were analysed by ANOVA (Statistica for Windows, Stat Soft Inc., Tulsa, OK, USA). No significant differences (P > 0.05) were found between DT of all passages. The DT was greater (P < 0.05) for BM (3.2 ± 1.5) than for AT (1.3 ± 0.7). By passage 8, AT MSC underwent 37.3 ± 4.6 cell-doublings (CD); by passage 5, BM MSC underwent 26.2 ± 5.03 CD. Positive von Kossa and Alizarin Red staining confirmed osteogenesis. Alcian blue staining illustrated chondrogenesis, and positive Oil Red O staining suggested adipogenesis. The AT and BM MSC were positive for CD90, CD44, and CD105; all cell lines were negative for haematopoietic markers such as CD34, CD14, and CD45. Although marker CD73 expresses reaction in other studies involving MSC in different species, it did not cross-react with equine AT and BM MSC. Results obtained revealed the immunophenotypic characterisation of the surface of isolated and cultivated MSC, classifying these cells as a promising type of progenitor cells that can be applied in equine cellular therapy.

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