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Objectives: To learn an easy and fast way to harvest lipoaspirate, isolate ADSC using an office based disposable system, and deploy these regenerative cells for different indications and anatomical regions.

Introduction: Adipose tissue derived stromal vascular fraction (SVF) is a reliable source of stromal vascular cells with regenerative potential in plastic surgery and other specialties. Although the gold standard, this enzymatic isolation method is expensive, requires expert personnel, raises legal and administrative concerns on the alteration of cell characteristics. We use a mech isolation system with comparable results

Materials / method: A total of 40 ml lipoaspirate was harvested from 60 women undergoing liposuction and was submitted to conventional enzymatic digestion for SVF isolation or mechanical digestion using a closed unit harnessing 3 ports with blades, followed by buffer incubation and centrifugation. Culture of the SVFs and flow cytometry were performed.

Results: The SVF cell yield obtained by enzymatic digestion was significantly higher 3,38 x 106/ml (±3.63, n=35) than that obtained by mechanical digestion 1,34 x 106/ml (±1.69, n=35), p=0,015. The average cell viability was 82.86% ±10.68 after enzymatic digestion versus 85,86% ±5.74 after mechanical digestion, which was not significant. Mechanical digested SVF expressed 2-fold higher stem cell surface markers compared with enzymatically digested SVF. Mechanical digestion was less time consuming, cost effective and didn't require a specific lab environment.

Conclusion: Mechanically digested SVF was comparable to enzymatically digested SVF in terms of stromal cell composition and viability. With mechanical digestion, we can isolate 30 to 50% SVF cells of that isolated with enzymatic digestion. Clinical outcomes seem very promising. Further studies are underway.

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Por favor indique a entidade (pessoa ou empresa): Minor shareholder of the device company

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Objectives: MEST: MEchanical SVF Transfer technique is the new way of producing SVF from fat tissue without enzyme in high amount of cells and viability. We are going to present our patented protocol for producing mechanical SVF with special device named as Adinizer. This device consisted of ultra sharp blade system and cuts the bond and bridges between adipocytes and mesechymal tissues under minimal pressure. Participants will learn how to mechanical SVF can be done with high amount of cells and viability with using Adinizer without causing to kill adipocytes.

Introduction: SVF can be isolated from adipose tissue either by means of enzymatic dissociation or by means of mechanical dissociation. Most enzymatic techniques used thus far for isolating the SVF from adipose tissue use collagenase. Main disadvantage of all these enzymatic techniques is that they are rather time consuming and expensive, but also that enzymatic treatment disrupts all communicative connections that exist between the cells as well as between the cells and ECM. Moreover, regulations in several countries does not allow to clinically apply cell-based products that are derived with collagenase.

Materials / method: 91 patients have been treated with Mechanical SVF Treatment (MEST) technique by using Adinizer in different indication. Patients have been followed minimum 6 months postoperatively. Cell count and viability have been evaluated by using dual florescence cell counter. Also our technique has been compared with enzyme technique in same patients in cGMP, cGLP laboratory with flow cytometry.

Results: All patients has been treated successfully with MEST technique. In flow flow cytometric analysis total cells were 2,5 X 10(6) viability was 96.68% in MEST group and respectively 3.5 X 10(6) and 98.23% in enzyme group. Also, cell count has been calculated up to 9.2 X 10(6) and viability up tom 99 % in dual florescence cell counter.

Conclusion: MEST might be more suitable as compared to enzymatic dissociation, because mechanical dissociation of adipose tissue is a fast time sparing inexpensive method and keeps the cell connections . Last 3 years there is shift form enzyme to mechanic SVF production. Since the adipose tissue is the most fragile tissue in the body, we believe that to get best results we should behave the fat tissue "gently".We developed and got patent a new approach as "Gentle Touch System". We cut the fat tissue with ultra sharp blades (Adinizer) to break bond and bridges between the adipocyte and mesenchymal cells .

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Por favor informe a data...: Completed
e o status: January 2017

Este trabalho não recebeu nenhum patrocínio direto ou indireto. O mesmo está sob a própria responsabilidade do seu autor.

Objectives: Adipose tissue is known a great source of cells for regenerative medicine therapeutic approaches. Since the invention of superiority of (freshly) isolated cells compared to conventional lipoinjection, the SVF injections are getting increased as the lipoinjection is offering a lower rate of viability due to partial necrosis of the grafted and transferred tissue.

Introduction: Stromal and Vascular Cells are a group of cells which are fixed and settled in the adipose tissue, are in the vascular walls and are adhered to the ExtraCellularMatrix (ECM). Usually, lipoaspiration procedure is performed to harvest the adipose tissue which Stromal and Vascular Cells are to be isolated from the tissue matrix. An alternative to enzyme use, we demonstrated here protocols of in-vitro study of cells dissociated using Microlyzer from manually harvested lipoaspirate.

Materials / method: Manually harvested lipoaspirate is used in this study. The design of Microlyzer offers 2-way connectors of Microlyzer bodies with luer-connectors for syringes. The Microlyzer body is a nest for the blade-filters. The blade-filters are 2400 to 600 microns. The device requires up to 3 steps of cutting of lipoaspirate for mechanical separation of the cells from the ECM. We used Luna Stem (Logos Bio, S. Korea) which can identify the population of nucleated and non-nucleated cells. Mechanical methods from literature offers wide range of nucleated cell counts/ml with different viability rates.

Results: Final product is roughly 3-5ml. SVF cells are meant to be counted focusing on nucleated cell population importantly. The results show that the Microlyzer method of SVF protocol offers wide range of remarkable nucleated cell counts up to 8,66 million nucleated cells/mL. Flow Cytometry (Beckman Coulter, USA) analyse is applied with promising results immunphenotypically, offering CD34, CD105, CD73, CD146 and CD90 cells were found remarkably with up to 99,9% of viability in the freshly isolated SVF using Non-Enzymatic SVF protocol of Microlyzer.

Conclusion: Further studies are needed to conclude the superiority of mechanical isolation of Stromal Vascular Fraction either safety or efficacy outcomes compared to the enzymatic isolation methods of Stromal Vascular Fraction.

These findings have shown that the Microlyzer device & Non-enzymatic SVF protocol offers a great yield capacity with higher viability in safe conditions as the protocol does not require any enzyme which offering risk-free of enzymatic activity within safety and regulation point of view.

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Por favor indique a entidade (pessoa ou empresa): T-LAB

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Por favor informe a data...: patent pending
e o status: 2018

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