Isolation of adipose tissue derived regenerative cells from human subcutaneous tissue with or without the use of an enzymatic reagent

Abstract

Background: Freshly isolated, unmodified autologous adipose derived regenerative cells (ADRCs) have emerged as a promising tool for regenerative cell therapy. The Transpose RT system (InGeneron, Inc., Houston, TX, USA) is a system for isolating ADRCs from adipose tissue, commercially available in Europe as a CE-marked medical device and under clinical evaluation in the United States. This system makes use of the proprietary, enzymatic Matrase Reagent for isolating cells. The present study addressed the question whether the use of Matrase Reagent influences cell yield, cell recovery, cell viability, biological characteristics, physiological functions or structural properties of the ADRCs in final cell suspension. Methods: Identical samples of subcutaneous adipose tissue from 12 subjects undergoing elective lipoplasty were processed either with or without the use of Matrase Reagent. Then, characteristics of the ADRCs in the respective final cell suspensions were evaluated. Results: Compared to non-enzymatic isolation, enzymatic isolation resulted in approximately twelve times higher mean cell yield (i.e., numbers of viable cells/ml lipoaspirate) and approximately 16 times more colony forming units (CFUs). Despite these differences, cells isolated from lipoaspirate both with and without the use of Matrase Reagent were independently able to differentiate into cells of all three germ layers. Discussion: The data of this study indicate that biological characteristics, physiological functions or structural properties relevant for the intended use were not altered or induced using Matrase Reagent. A comprehensive literature review demonstrated that isolation of ADRCs from lipoaspirate using the Transpose RT system and the Matrase Reagent results in the highest viable cell yield among all published data regarding isolation of ADRCs from lipoaspirate. ABSTRACTBackground: Freshly isolated, unmodified autologous adipose derived regenerative cells (ADRCs) have emerged as a promising tool for regenerative cell therapy. The Transpose RT system (InGeneron, Inc., Houston, TX, USA) is a system for isolating ADRCs from adipose tissue, commercially available in Europe as a CE-marked medical device and under clinical evaluation in the United States. This system makes use of the proprietary, enzymatic Matrase Reagent for isolating cells. The present study addressed the question whether the use of Matrase Reagent influences cell yield, cell recovery, cell viability, biological characteristics, physiological functions or structural properties of the ADRCs in final cell suspension. Methods: Identical samples of subcutaneous adipose tissue from 12 subjects undergoing elective lipoplasty were processed either with or without the use of Matrase Reagent. Then, characteristics of the ADRCs in the respective final cell suspensions were evaluated. Results: Compared to non-enzymatic isolation, enzymatic isolation resulted in approximately twelve times higher mean cell yield (i.e., numbers of viable cells/ml lipoaspirate) and approximately 16 tim...

Unmodified autologous stem cells at point of care for chronic myocardial infarction

“…A number of enzymatic and non-enzymatic systems for isolating UA-ADRCs have been developed[21,54]. The cell yield reported after different procedures varies considerably[21,55].…”

“…A number of enzymatic and non-enzymatic systems for isolating UA-ADRCs have been developed[21,54]. The cell yield reported after different procedures varies considerably[21,55]. It was shown that, in general, enzymatic isolation of UA-ADRCs yields significantly more cells than non-enzymatic, mere mechanical isolation[21,56].…”

“…To this end, each aliquot was minced and incubated together with enzymatic Matrase Reagent (InGeneron) for 1 h under agitation in the processing unit at 39°C, according to the manufacturer’s instructions for use. Note that UA-ADRCs that were isolated from adipose tissue with the Transpose RT system and the enzymatic Matrase Reagent were comprehensively characterized in a number of studies[19-21]. This included the demonstrated expression of the regenerative cell-associated genes Oct4, Klf4 and Hes3[21], as well as their differentiative potential into adipogenic, osteogenic, hepatogenic and neurogenic cell lines[21].…”

“…Note that UA-ADRCs that were isolated from adipose tissue with the Transpose RT system and the enzymatic Matrase Reagent were comprehensively characterized in a number of studies[19-21]. This included the demonstrated expression of the regenerative cell-associated genes Oct4, Klf4 and Hes3[21], as well as their differentiative potential into adipogenic, osteogenic, hepatogenic and neurogenic cell lines[21]. On this basis, UA-ADRCs isolated from adipose tissue with the Transpose RT system and the enzymatic Matrase Reagent were used in a number of clinical pilot trials[22-24], and are currently under investigation in a number of Investigational Device Exemption (IDE) studies approved by the United States Food and Drug Administration[25-29].…”

Effect of Hypoxia Preconditioned Secretomes on Lymphangiogenic and Angiogenic Sprouting: An in Vitro Analysis

“…ADCS is known for its pro-angiogenic and lymphangiogenic activity [ 31 , 32 , 33 , 34 ], therefore it was selected as a positive control for this study. According to manufacturer’s (ARC Processing System, InGeneron Inc., Houston, TX, USA) instructions and a previously published protocol [ 35 ], adipose tissue was dissected from female adult mice, (approximately 10–12 g) using appropriate surgical techniques into a sterile container and then manually minced until average piece size was approximately 2 mm or less. The minced adipose tissue was transferred into a sterile tube and lactated ringer (preheated to 39 °C) was added to a fill level of 30 mL.…”