A Proliferation and surface marker characterization of adipose stem cells after culture in various processed outdated platelet lysate containing media

Sari, Puji; Luviah, Evah; Purwoko, Reza Yuridian; Pawitan, Jeanne Adiwinata

doi:10.20902/ijptr.2017.10118

Cited by 2 publications

(2 citation statements)

References 28 publications

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“…Platelet-rich plasma (PRP) has high growth factors content [6,7]. In in vitro studies, PRP was used as the source of growth factors to support MSC proliferation and differentiation [6,[22][23][24][25], replacing the use of animal-derived serum. In animal and human studies, PRP was used for accelerating wound healing process [26], improving regeneration of injured articular cartilage [6,7], revising hypertrophic scars [21], and reducing the necrosis that frequently occurred in fat graft procedure [27].…”

Section: Introductionmentioning

confidence: 99%

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

Karina¹,

Rosliana²,

Rosadi³

et al. 2020

Scientifica

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Background. Stromal vascular fraction (SVF) therapy has been performed over the past six years to treat 421 patients by our group in five clinical centers. Autologous SVF, which is a substance containing stem cells, was isolated from lipoaspirate, mixed with platelet-rich plasma (PRP), and administered to patients with degenerative diseases, autoimmune diseases, trauma, aging, and other diseases with unknown etiology. This study aimed to determine the safety of SVF and PRP that were given through infusion, spinal, and intra-articular injection. Methods. The lipoaspirate was treated with a tissue-dissociating enzyme, and then, through centrifugation, SVF was isolated. In addition, blood was drawn from each patient, and PRP was isolated. Autologous PRP and SVF were administered to all subjects by intravenous (IV) injection. A minority group within the population received an additional spinal or intra-articular injection. The type of intervention was determined by each disease evaluation. The cell doses and adverse events for each patient were documented and analyzed. Results. Cell dose that was considered to be safe was less than 10 billion SVF cells in 250 cc of normal saline, for IV injection, and less than 1 billion SVF, for intra-articular and spinal injection. Adverse events were not severe and were treated successfully. Any observed adverse events were identified as a result of spinal or intra-articular injections and were not related to SVF or PRP. Conclusions. Our results showed that administration of high dose of SVF until 10 billion cells in a majority of 421 patients through infusion, spinal, and intra-articular injection was feasible without causing major adverse events and should be further investigated in well-designed phase I-II clinical trial to address the safety and efficacy of therapy.

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Section: Introductionmentioning

confidence: 99%

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

Karina¹,

Rosliana²,

Rosadi³

et al. 2020

Scientifica

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“…Outdated hPL showed highest concentration of TGF-β1 compared to other growth factors, 20 and TGF-β1 level was not influences by freeze-thaw cycles. 21 This fact might influence the expression of CD73 on Ps-3, which had low expression (below 80%).…”

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confidence: 78%

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2017

IJPSR

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Human adipose tissue-derived stem cells (hADSCs) are promising for cellular therapy as they are multipotent, which consistently express high CD90, CD105, CD73 and low hematopoietic surface markers. Flow cytometry is a routine technology that is used in most laboratories and can be used to study hADSC characterization, but the instrument needs strict calibration to assure the quality of the results. However, for rarely used fluorophores, calibration is often neglected. This study aimed to compare calibrated and non-calibrated fluorescence (FL) in flow cytometry performances. hADSCs were isolated from lipoaspirate and cultured in human platelet lysate supplemented medium. Passaged cells were used for characterization. Cells stained with positive (CD90-FITC, CD105-PerCP Cy5.5, CD73-APC) and negative (CD34, CD45, CD11b, CD19, HLA-DR)-PE marker cocktails were subjected to flow cytometry analysis. In the instrument, FITC, PE, PerCP-Cy5.5, and APC were detected by FL1, FL2, FL3, and FL4 respectively. Routine calibration of FL1, FL2, and FL3 was done, except for FL4. Therefore, data from calibrated and non-calibrated FL4 were analyzed. hADSCs strongly and consistently expressed CD90, CD105, and negative surface markers at averages of 94.6%, 90.5%, and 2.8% respectively, hence in line with ISCT guidelines; in contrast to CD73 (FL4), whose average was 42%. Characterization of calibrated FL4 showed very good results on CD73 (above 94%), but not in non calibrated FL4, which showed inconsistent results, as values ranged between 0% -82%. In conclusion, routine calibration of FL1, FL2, FL3, and FL4 is important to get a reliable result.

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A Proliferation and surface marker characterization of adipose stem cells after culture in various processed outdated platelet lysate containing media

Cited by 2 publications

References 28 publications

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

Safety of Technique and Procedure of Stromal Vascular Fraction Therapy: From Liposuction to Cell Administration

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