Does the Harvesting Technique Affect the Properties of Adipose‐Derived Stem Cells?—The Comparative Biological Characterization

Bajek, Anna; Gurtowska, Natalia; Olkowska, Joanna; Maj, Małgorzata; Kaźmierski, Łukasz; Bodnar, Magdalena; Marszałek, Andrzej; Dębski, Robert; Drewa, Tomasz

doi:10.1002/jcb.25724

Cited by 36 publications

(40 citation statements)

References 51 publications

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“…Earlier studies reported the heterogeneity in SVF and great variability in the percentages of different subsets [ 35 , 36 ]. Some reports have focused mostly on the difference in the isolation methods, handling, variability of cell yields and mode of administration [ 37 , 38 ]. To date, there are insufficient data to establish a correlation between SVF-dose, and direct therapeutic effect.…”

Section: Resultsmentioning

confidence: 99%

The ratio of ADSCs to HSC‐progenitors in adipose tissue derived SVF may provide the key to predict the outcome of stem‐cell therapy

Kilinc¹,

Santidrián²,

Minev³

et al. 2018

Clinical & Translational Med

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Background Stromal vascular fraction (SVF) represents an attractive source of adult stem cells and progenitors, holding great promise for numerous cell therapy approaches. In 2017, it was reported that 1524 patients received autologous SVF following the enzymatic digestion of liposuction fat. The treatment was safe and effective and patients showed significant clinical improvement. In a collaborative study, we analyzed SVF obtained from 58 patients having degenerative, inflammatory, autoimmune diseases, and advanced stage cancer. Results Flow analysis showed that freshly isolated SVF was very heterogeneous and harbored four major subsets specific to adipose tissue; CD34 high CD45 − CD31 − CD146 − adipose-derived stromal/stem cells (ADSCs), CD34 low CD45 + CD206 + CD31 − CD146 − hematopoietic stem cell-progenitors (HSC-progenitors), CD34 high CD45 − CD31 + CD146 + adipose tissue-endothelial cells and CD45 − CD34 − CD31 − CD146 + pericytes. Culturing and expanding of SVF revealed a homogenous population lacking hematopoietic lineage markers CD45 and CD34, but were positive for CD90, CD73, CD105, and CD44. Flow cytometry sorting of viable individual subpopulations revealed that ADSCs had the capacity to grow in adherent culture. The identity of the expanded cells as mesenchymal stem cells (MSCs) was further confirmed based on their differentiation into adipogenic and osteogenic lineages. To identify the potential factors, which may determine the beneficial outcome of treatment, we followed 44 patients post-SVF treatment. The gender, age, clinical condition, certain SVF-dose and route of injection, did not play a role on the clinical outcome. Interestingly, SVF yield seemed to be affected by patient’s characteristic to various extents. Furthermore, the therapy with adipose-derived and expanded-mesenchymal stem cells (ADE-MSCs) on a limited number of patients, did not suggest increased efficacies compared to SVF treatment. Therefore, we tested the hypothesis that a certain combination, rather than individual subset of cells may play a role in determining the treatment efficacy and found that the combination of ADSCs to HSC-progenitor cells can be correlated with overall treatment efficacy. Conclusions We found that a 2:1 ratio of ADSCs to HSC-progenitors seems to be the key for a successful cell therapy. These findings open the way to future rational design of new treatment regimens for individuals by adjusting the cell ratio before the treatment.

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Section: Resultsmentioning

confidence: 99%

The ratio of ADSCs to HSC‐progenitors in adipose tissue derived SVF may provide the key to predict the outcome of stem‐cell therapy

Kilinc¹,

Santidrián²,

Minev³

et al. 2018

Clinical & Translational Med

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“…The content of preadipocytes is less in the primary ADSCs 'mix' (23,25,26) and preadipocytes share several cell markers with the primary adipose derived stem cells, like Lin -, CD29 + , CD34 + , Sca-1 + , and CD45 -CD31 - (23,25), making it difficult to screen for preadipocytes alone from the primary ADSC 'mix' using fluorescence-activated cell sorting. In the clinical practice of regenerative medicine, generally, the ADSCs was used and observed as a whole (3)(4)(5)(6).…”

Section: Discussionmentioning

confidence: 99%

“…Culturing with increasing cell passages was found to reduce the heterogeneity of ADSCs (11). Studies indicated that continuous cell passages reduced the number of cells with a CD34 + surface marker, which differentiated into adipocytes more efficiently (25,27). However, whether spontaneous adipogenic differentiation of ADSCs would decrease with serial cell passages is not yet clear.…”

Section: Discussionmentioning

confidence: 99%

Spontaneous adipogenic differentiation potential of adipose‑derived stem cells decreased with increasing cell passages

Yang

Zhang

2018

Mol Med Report

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Primary adipose-derived stem cells (ADSCs) are a mixture of cell types including preadipocytes having the ability to spontaneously differentiate into adipocytes. The aim of the present study was to compare the spontaneous adipogenic differentiation potential of ADSCs at different passages to determine whether it decreased with continuous cell passages. Mouse ADSCs (mADSCs) were harvested and cells from passages 1 to 5 were used for experiments. The proliferation of mADSCs at different passages was tested using the cell counting kit‑8 assay. Reverse transcription‑quantitative polymerase chain reaction was used to determine relative mRNA expression levels of CCAAT/enhancer binding protein α (Cebpa and C/EBPα) and peroxisome proliferator‑activated receptor γ (Pparg and PPARγ), and western blot analysis was used to investigate C/EBPα and PPARγ protein expression. The cells were cultured using DMEM. The fixed cells were then stained using Oil Red O on days 14 and 28, and the obtained extracted dye was monitored for absorbance. The 510 nm absorbance from passages 1 to 5 was observed to be statistically different. The relative expression levels of Cebpa and Pparg for mADSCs from passage 1 were significantly higher when compared with those for mADSCs from passages 2 to 5 on days 3, 5 and 7. However, no difference was identified in the expression levels of proteins C/EBPα and PPARγ for different passages. Although the mRNA expression levels of Pparg from passages 4 to 5 were significantly higher when compared with those from passage 1, the results of Oil Red O absorbance, mRNA expression levels of Cebpa, and the protein expression levels of C/EBPα and PPARγ exhibited no difference between mADSCs from passages 1 to 5 when cultured with induced adipogenic differentiation medium. Therefore, it was concluded that the spontaneous adipogenic differentiation potential of ADSCs decreased with continuous cell passages.

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“…Differentiation capacity into the adipogenic, osteogenic, and chondrogenic lineage was performed as described previously [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

“…Stem cells were additionally analyzed for the presence of the specific surface markers CD34, CD44, CD45, and CD90 by flow cytometry according to the protocol previously described [ 21 ].…”

Section: Methodsmentioning

confidence: 99%

Human Adipose-Derived and Amniotic Fluid-Derived Stem Cells: A Preliminary In Vitro Study Comparing Myogenic Differentiation Capability

et al. 2018

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BackgroundAround the world, disabilities due to musculoskeletal disorders have increased and are a major health problem worldwide. In recent years, stem cells have been considered to be powerful tools for musculoskeletal tissue engineering. Human adipose-derived stem cells (hADSCs) and amniotic fluid-derived stem cells (hAFSCs) undergo typical differentiation process into cells of mesodermal origin and can be used to treat muscular system diseases. The aim of the present study was to compare the biological characteristic of stem cells isolated from different human tissues (adipose tissue and amniotic fluid) with respect to myogenic capacity and skeletal and smooth muscle differentiation under the same conditions.Material/MethodshAFSCs and hADSCs were isolated during standard medical procedures and widely characterized by specific markers expression and differentiation potential. Both cell types were induced toward smooth and striated muscles differentiation, which was assessed with the use of molecular techniques.ResultsFor phenotypic characterization, both stem cell types were assessed for the expression of OCT-4, SOX2, CD34, CD44, CD45, and CD90. Muscle-specific markers appeared in both stem cell types, but the proportion of positive cells showed differences depending on the experimental conditions used and the source from which the stem cells were isolated.ConclusionsIn this study, we demonstrated that hADSCs and hAFSCs have different capability of differentiation toward both muscle types. However, hADSCs seem to be a better source for myogenic protocols and can promote skeletal and smooth muscle regeneration through either direct muscle differentiation or by paracrine mechanism.

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Does the Harvesting Technique Affect the Properties of Adipose‐Derived Stem Cells?—The Comparative Biological Characterization

Cited by 36 publications

References 51 publications

The ratio of ADSCs to HSC‐progenitors in adipose tissue derived SVF may provide the key to predict the outcome of stem‐cell therapy

The ratio of ADSCs to HSC‐progenitors in adipose tissue derived SVF may provide the key to predict the outcome of stem‐cell therapy

Spontaneous adipogenic differentiation potential of adipose‑derived stem cells decreased with increasing cell passages

Human Adipose-Derived and Amniotic Fluid-Derived Stem Cells: A Preliminary In Vitro Study Comparing Myogenic Differentiation Capability

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