scholarly journals Cryopreservation Increases Availability and Usability of Ovine Adipose-tissue Derived Stem Cells for Regenerative Medicine Search

2001 ◽  
Vol 71 (3) ◽  
pp. 178-184
Author(s):  
Ionela Movileanu ◽  
Klara Brinzaniuc ◽  
Marius M. Harpa ◽  
Ovidiu S. Cotoi ◽  
Terezia Preda ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Cell Loss ◽  
Blue Staining ◽  
Minimal Cell ◽  
Differentiation Potential ◽  
Preservation Method

Aim: Our long-term aim is to develop a living valvular substitute using Regenerative Medicine principles, by seeding decellularized porcine heart valve scaffolds with adult stem cells and conditioning them in bioreactors before implantation. In this study, adult stem cells were isolated from sheep adipose tissue (ADSCs). However, we found it impractical to use cells immediately after propagation and thus, in order to extend their availability in time, a preservation method was needed. Methods: Adipose tissue was harvested from 6 sheep. ADSCs were isolated using enzymatic agents and cultured. The cells were tested for plasticity using chondrogenic, adipogenic and osteogenic differentiation kits and then cryopreserved in DMSO at -1400C. Viability was tested after a 3 week storage using Trypan Blue Staining. Results: Ovine ADSCs exhibited excellent plasticity and differentiation potential. An average of 18 million ADSCs were obtained from each ovine, exhibiting more than 88% viability after a 3-week cryopreservation period followed by thawing. Conclusions: DSMO cryopreservation represents a suitable method for ovine ADSCs for regenerative medicine. This method expands the usage of stem cells in vitro before they are differentiated into more specialized cells, offering large numbers of usable ADSCs with minimal cell loss at any desired time point.

scholarly journals Fundamentals of Stem Cells: Why and How Patients' Own Adult Stem Cells Are the Next Generation of Medicine

2019 ◽  
Author(s):  
Eckhard U. Alt ◽  
Christoph Schmitz ◽  
Xiaowen Bai
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Stem Cell ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Point Of Care ◽  
Appropriate Technology ◽  
Differentiation Potential ◽  
Resident Stem Cells ◽  
New Generation

Various tissue resident stem cells are receiving attention from basic scientists and clinicians as they hold certain promise for regenerative medicine. This paper is intended to clarify and facilitate the understanding, development and adoption of regenerative medicine in general and specifically of therapies based on unmodified, autologous adipose-derived regenerative cells (UA-ADRCs). To this end, results of landmark experiments on stem cells and stem cell therapy performed in the labs of the authors are summarized, the most intriguing of which are the following: (i) vascular associated mesenchymal stem cells (MSCs) can be isolated from different organs (adipose tissue, heart, skin, bone marrow and skeletal muscle) and differentiated into ectoderm, mesoderm and endoderm, providing significant support for the hypothesis of the existence of a small, ubiquitously distributed, universal vascular associated stem cell with full pluripotency; (ii) the orientation and differentiation of MSCs are driven by signals of the respective microenvironment; and (iii) these stem cells irrespective of the tissue origin exhibit full pluripotent differentiation potential without any prior genetic modification or the need for culturing. They can be obtained from a small amount of adipose tissue when using the appropriate technology for isolating the cells, and can be harvested from and re-applied to the same patient at the point of care without the need for complicated processing, manipulation, culturing, expensive equipment, or repeat interventions. These findings demonstrate the potential of UA-ADRCs for triggering the development of an entire new generation of medicine for the benefit of patients and of healthcare systems.

scholarly journals Adipose-Derived Stem Cells for Future Regenerative System Medicine

2012 ◽  
Vol 4 (2) ◽  
pp. 59
Author(s):  
Yani Lina ◽  
Andi Wijaya
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Adipose Tissue ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Stromal Vascular Fraction ◽  
Human Adipose Tissue ◽  
Lytic Enzymes ◽  
Stromal Stem Cells

BACKGROUND: The potential use of stem cell-based therapies for repair and regeneration of various tissues and organs offers a paradigm shift that may provide alternative therapeutic solutions for a number of disease. Despite the advances, the availability of stem cells remaining a challenge for both scientist and clinicians in pursuing regenerative medicine. CONTENT: Subcutaneous human adipose tissue is an abundant and accessible cell source for applications in tissue engineering and regenerative medicine. Routinely, the adipose issue is digested with collagenase or related lytic enzymes to release a heterogeneous population for stromal vascular fraction (SVF) cells. The SVF cells can be used directly or can be cultured in plastic ware for selection and expansion of an adherent population known as adipose-derived stromal/stem cells (ASCs). Their potential in the ability to differentiate into adipogenic, osteogenic, chondrogenic and other mesenchymal lineages, as well in their other clinically useful properties, includes stimulation of angiogenesis and suppression of inflammation.SUMMARY: Adipose tissue is now recognized as an accessible, abundant and reliable site for the isolation of adult stem cels suitable for the application of tissue engineering and regenerative medicine applications. The past decade has witnessed an explosion of preclinical data relating to the isolation, characterization, cryopreservation, differentiation, and transplantation of freshly isolated stromal vascular fraction cells and adherent, culture-expanded, adipose-derived stromal/stem cells in vitro and in animal models.KEYWORDS: adipose tissue, adult stem cells, regenerative medicine, mesenchymal stem cells

scholarly journals Sources and Clinical Applications of Mesenchymal Stem Cells: State-of-the-art review

2018 ◽  
Vol 18 (3) ◽  
pp. 264 ◽  
Author(s):  
Roberto Berebichez-Fridman ◽  
Pablo R. Montero-Olvera
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Current Knowledge ◽  
Clinical Applications ◽  
The Body ◽  
Differentiation Potential ◽  
Advantages And Disadvantages

First discovered by Friedenstein in 1976, mesenchymal stem cells (MSCs) are adult stem cells found throughout the body that share a fixed set of characteristics. Discovered initially in the bone marrow, this cell source is considered the gold standard for clinical research, although various other sources—including adipose tissue, dental pulp, mobilised peripheral blood and birth-derived tissues—have since been identified. Although similar, MSCs derived from different sources possess distinct characteristics, advantages and disadvantages, including their differentiation potential and proliferation capacity, which influence their applicability. Hence, they may be used for specific clinical applications in the fields of regenerative medicine and tissue engineering. This review article summarises current knowledge regarding the various sources, characteristics and therapeutic applications of MSCs.Keywords: Mesenchymal Stem Cells; Adult Stem Cells; Regenerative Medicine; Cell Differentiation; Tissue Engineering.

scholarly journals Application of Nanoscaffolds in Mesenchymal Stem Cell-Based Therapy

2014 ◽  
Vol 2014 ◽  
pp. 1-14 ◽  
Author(s):  
Saman Ghoraishizadeh ◽  
Afsoon Ghorishizadeh ◽  
Peyman Ghoraishizadeh ◽  
Nasibeh Daneshvar ◽  
Mohadese Hashem Boroojerdi
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Regenerative Medicine ◽  
Adult Stem Cells ◽  
Cell Signalling ◽  
Great Influence ◽  
Source Characterization ◽  
Differentiation Potential ◽  
Cell Based Therapy ◽  
Essential Components

Regenerative medicine is an alternative solution for organ transplantation. Stem cells and nanoscaffolds are two essential components in regenerative medicine. Mesenchymal stem cells (MSCs) are considered as primary adult stem cells with high proliferation capacity, wide differentiation potential, and immunosuppression properties which make them unique for regenerative medicine and cell therapy. Scaffolds are engineered nanofibers that provide suitable microenvironment for cell signalling which has a great influence on cell proliferation, differentiation, and biology. Recently, application of scaffolds and MSCs is being utilized in obtaining more homogenous population of MSCs with higher cell proliferation rate and greater differentiation potential, which are crucial factors in regenerative medicine. In this review, the definition, biology, source, characterization, and isolation of MSCs and current report of application of nanofibers in regenerative medicine in different lesions are discussed.

3D printable Sodium alginate-Matrigel (SA-MA) hydrogel facilitated ectomesenchymal stem cells (EMSCs) neuron differentiation

2020 ◽  
Vol 35 (6) ◽  
pp. 709-719 ◽  
Author(s):  
Yang Li ◽  
Xia Cao ◽  
Wenwen Deng ◽  
Qingtong Yu ◽  
Congyong Sun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Sodium Alginate ◽  
Neuronal Differentiation ◽  
Adult Stem Cells ◽  
Three Dimensional ◽  
Cell Types ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Cord Injury

Ectomesenchymal stem cells (EMSCs) are typical adult stem cells obtained from the cranial neural crest. They have the potential to differentiate into various cell types, such as osseous cells, neurons and glial cells. Three-dimensional (3 D) printing is a novel method to construct biological structures by rapid prototyping. Previously, our group reported on the stemness and multi-lineage differentiation potential of EMSCs on gels. However, the exploration of EMSCs in 3 D printing and then evaluation of the growth and neuronal differentiation of EMSCs on extruded 3 D printable hybrid hydrogels has not been reported. Therefore, the current study explored the novel hybrid Sodium alginate-Matrigel (SA-MA) hydrogel extruded 3 D printing to design an in vitro scaffold to promote the differentiation and growth of EMSCs. In addition, the physical properties of the hydrogel were characterized and its drug-releasing property determined. Notably, the results showed that the construct exhibited a sustain-released effect of growth factor BDNF in accordance with the Higuchi equation. Moreover, the cell survival rate on the 3 D printed scaffold was 88.22 ± 1.13% with higher neuronal differentiation efficiency compared with 2 D culture. Thus, SA-MA’s ability to enhanced EMSCs neuronal differentiation offers a new biomaterial for neurons regeneration in the treatment of spinal cord injury.

202 IN VITRO DIFFERENTIATION OF ADULT PORCINE ADIPOSE-DERIVED STEM CELLS AFTER LABELING WITH PKH26 AND FLOW CYTOMETRY

2006 ◽  
Vol 18 (2) ◽  
pp. 209
Author(s):  
M. Mello ◽  
A. Lima ◽  
S. Malusky ◽  
S. Lane ◽  
M. Wheeler
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Adipose Tissue ◽  
Osteogenic Differentiation ◽  
Fluorescent Dye ◽  
Positive Staining ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential ◽  
Oil Red O

The purpose of this study was to investigate the possible effects of the fluorescent dye PKH26 and flow cytometry on adult porcine adipose-derived stem cells (ADSCs) after exposing them to adipogenic and osteogenic differentiation conditions. Adipose tissue was isolated from swine (11 months of age) and digested with 0.075% collagenase at 37�C for 90 min. The digested adipose tissue was centrifuged at 200g for 10 min to obtain a cell pellet. The pellet was re-suspended with DMEM, and the ADSCs were plated onto 75 cm2 flasks (5000-10 000 cells per cm2) and cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and 1% gentamicin. Passage 3 cells were labeled with fluorescent dye (PKH26 red fluorescent cell linker kit; Sigma Chemical, St. Louis, MO, USA) and sorted by flow cytometry. After labeling and sorting, the sorted and unsorted (control group) cells were replated and exposed to adipogenic (1 �M dexamethasone, 0.5 mM isobutylmethylxantine, 10 �M insulin and 200�M indomethacin) and osteogenic (0.1 �M dexamethasone, 10 mM �-glycerophosphate, and 50�M ascorbic acid) differentiation conditions when the cells were 90% confluent. Cells were evaluated based on morphology and specific staining properties. Adipogenic differentiation was confirmed by oil red O-positive staining of large lipid vacuoles, and osteogenic differentiation by Von Kossa staining 2 weeks after initiation of differentiation. The frequency of oil red O-positive colonies in both sorted and unsorted group was similar (15.0% vs. 13.2%, respectively). The number of osteogenic nodules, confirmed by the presence of calcium by Von Kossa staining, in the sorted and unsorted group was 17 and 184 per flask, respectively. In conclusion, this study demonstrates that adult porcine adipose-derived stem cells maintain their differentiation potential after labeling with fluorescent dye and sorting by flow cytometry. This should allow for more rapid evaluation of the differentiation potential of ADSCs in vitro. This work was partially supported by the Council for Food and Agricultural Research (C-FAR) Sentinel Program, University of Illinois and CNPq, Brazil (M. Mello).

401 MIGRATION AND THERAPEUTIC POTENTIAL OF PORCINE ADULT ADIPOSE-DERIVED MESENCHYMAL STEM CELLS

2010 ◽  
Vol 22 (1) ◽  
pp. 357 ◽  
Author(s):  
S. M. Wilson ◽  
E. Monaco ◽  
M. S. Goldwasser ◽  
S. G. Clark ◽  
W. L. Hurley ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Subcutaneous Adipose Tissue ◽  
Adult Stem Cells ◽  
Bone Defects ◽  
Adipose Derived Stem Cells ◽  
Subcutaneous Adipose ◽  
Time Point

Bone marrow is one current source of adult stem cells for therapeutic purposes; however, the magnitude and accessibility of subcutaneous adipose tissue in humans make it an attractive alternative. Numerous in vitro studies have been conducted to determine how these cells act in vitro, but it is imperative to determine the vast abilities of these cells in vivo. The objective of this study was to evaluate in vivo migration and bone healing ability after transplanting adipose-derived stem cells (ADSC) in a swine model. Adipose-derived stem cells were isolated from subcutaneous adipose tissue of adult Yorkshire pigs and cultured in vitro. At 80 to 90% confluence/passage 3, the cells were trypsinized and labeled in suspension with carboxyfluorescein succinimidyl ester (CFDA-SE). This project included 20 pigs weighing between 63.5 and 81.7 kg. Bilateral mandibular osteoectomies with 10-mm defects were performed on each pig. Of the 20 pigs, half received a treatment of 2.5 million CFDA-SE labeled stem cells administered directly into each defect (DI), and the remaining half received a treatment of approximately 5 million CFDA-SE labeled stem cells through an ear vein injection via catheter (EVI). The time points were 1 h and 2 and 4 wk, with 2 pigs per time with the DI and EVI treatments. Pigs were slaughtered at each time, and spleen, liver, lung, kidney, ear vein, blood, and mandible tissues were collected. Blood samples were collected from the jugular vein with EDTA and processed via flow cytometry after collection. Tissues were fixed in 10% buffered formalin for histology. Fluorescent microscopy (CFDA-SE excitation/emission is 492/517 nm) has confirmed that transplanted ADSC do indeed migrate to a site of injury or trauma. Labeled cells were also present in blood collected from the 1-h time point group. Currently, we have not seen the presence of labeled ADSC in the other tissues (spleen, liver, lung, and kidney) after the 1-h time point. We did observe that ADSC administered by DI and EVI were able to significantly heal and regenerate bone defects within 4 wk post-surgery (P < 0.05, ANOVA with F-test), in contrast to bone defects in pigs that did not receive cell injections (control). Evidence of ADSC-related healing and bone regeneration was evident by gross visualization, dual-energy x-ray absorptiometry (DXA) and micro computer tomography (microCT) analysis. The clinical implications of these results are significant for treating many diseases in which inflammation or defects exist, such as cardiac disease, neurological disease, or traumatic injuries to both soft and hard tissue. If the adult stem cells can be harvested from fat, encouraged to produce bone or cartilage, and then reinserted into defects, treatment protocols for trauma victims could be developed that would reduce the need for alternate harvesting techniques for bone. This work was support by a grant from the Illinois Regenerative Medicine Institute (IDPH # 63080017).

381 LABELING AND ANALYSIS OF SWINE ADIPOSE-DERIVED STEM CELLS WITH CARBOXYFLUORESCEIN DIACETATE AND QUANTUM DOTS

2010 ◽  
Vol 22 (1) ◽  
pp. 347
Author(s):  
N. Cieslak ◽  
A. Massie ◽  
S. M. Wilson ◽  
E. Monaco ◽  
M. B. Wheeler
Keyword(s):  
Stem Cells ◽  
Quantum Dots ◽  
Regenerative Medicine ◽  
Quantum Dot ◽  
In Vitro Culture ◽  
Adult Stem Cells ◽  
Attractive Alternative ◽  
Adipose Derived Stem Cells

The quantity, accessibility, and abundance of subcutaneous adipose tissue in humans make it an attractive alternative to bone marrow as a source of adult stem cells for therapeutic purposes. Adult adipose-derived mesenchymal stem cells can differentiate into a variety of lineages including adipose, bone, cartilage, and muscle. In addition, the use of adult stem cells for regenerative medicine rather than those from embryos avoids concerns with ethics, safety, and immunology. One important issue is the ability to track the transplanted stem cells during the regeneration process to evaluate the stem cell-mediated healing. The objective of this study was to compare the efficiency, longevity, and intensity of carboxyfluorescein diacetate, succinimidyl ester (CFDA SE) and quantum dot nanocrystal (Qtracker™, Invitrogen, Carlsbad, CA, USA) labeled adipose-derived stem cells (ADSC) over an in vitro culture period of 4 weeks. Adipose-derived stem cells (6 x 106) previously isolated and frozen at -196°C were thawed and cultured in 75-cm3 flasks with 14 mL of DMEM. Cells were grown to 80% confluence and trypsinized. After trypsinization, the cells were divided into 4 treatments (3 x 106 cells per treatment). The treatments were (1) unlabeled control, (2) labeled with 30 μM CFDA SE, (3) labeled with 15 nM Qtracker™, and (4) labeled with 15 nM Qtracker™, following the Invitrogen Qtracker™ protocol. Cells (1 x 106) were removed from each treatment every week for 4 weeks and fixed in formalin for later analysis. When all the samples were collected, they were analyzed using flow cytometry. Data were analyzed via chi-square test. The percentage of cells labeled with CFDA SE and Qtracker™ was 99.35 and 98.46%, respectively, immediately after labeling. By 1 wk, the percentage of cells labeled with CFDA SE and Qtracker™ had deceased (P < 0.01) to 0.11 and 1.48%, respectively. The CFDA SE-labeled cell percentages had decreased (P < 0.01) to 0% at 2, 3, and 4 wk, respectively. The Qtracker™-labeled cells also decreased (P < 0.01) to 0.745, 1.69 and 0.45% at 2, 3, and 4 wk, respectively. The high rate of cell division of these cells in vitro might be responsible for the rapid loss of both labels during the first week of culture. Previous results from our lab have shown that the CFDA SE is retained in the cells for up to 6 wk in vivo (Lima AS et al. 2006 Reprod. Fertil. Dev. 18, 208). Similar studies need to be done with the quantum dot-labeled cells to determine the Qtracker™ label’s longevity in vivo. In conclusion, quantum dots can be used to label ADSC, in vitro, for at least 4 wk, albeit at much lower levels than those observed during the week following labeling. Determination of a suitable label for high-percentage porcine ADSC labeling during long-term in vitro culture remains to be completed. This research was supported by the Intel Scholar’s Program and the Illinois Regenerative Medicine Institute.

Human adipose tissue stem cells: relevance in the pathophysiology of obesity and metabolic diseases and therapeutic applications

2012 ◽  
Vol 14 ◽  
Author(s):  
Angelo Cignarelli ◽  
Sebastio Perrini ◽  
Romina Ficarella ◽  
Alessandro Peschechera ◽  
Pasquale Nigro ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Adult Stem Cells ◽  
Metabolic Diseases ◽  
Human Adipose Tissue ◽  
Cell Types ◽  
Differentiation Potential ◽  
Multipotent Stem Cells ◽  
Therapeutic Applications ◽  
Metabolic Characteristics

Stem cells are unique cells exhibiting self-renewing properties and the potential to differentiate into multiple specialised cell types. Totipotent or pluripotent stem cells are generally abundant in embryonic or fetal tissues, but the use of discarded embryos as sources of these cells raises challenging ethical problems. Adult stem cells can also differentiate into a wide variety of cell types. In particular, adult adipose tissue contains a pool of abundant and accessible multipotent stem cells, designated as adipose-derived stem cells (ASCs), that are able to replicate as undifferentiated cells, to develop as mature adipocytes and to differentiate into multiple other cell types along the mesenchymal lineage, including chondrocytes, myocytes and osteocytes, and also into cells of endodermal and neuroectodermal origin, including beta-cells and neurons, respectively. An impairment in the differentiation potential and biological functions of ASCs may contribute to the development of obesity and related comorbidities. In this review, we summarise different aspects of the ASCs with special reference to the isolation and characterisation of these cell populations, their relation to the biochemical features of the adipose tissue depot of origin and to the metabolic characteristics of the donor subject and discuss some prospective therapeutic applications.

scholarly journals Towards a Comprehensive Understanding of UA-ADRCs (Uncultured, Autologous, Fresh, Unmodified, Adipose Derived Regenerative Cells, Isolated at Point of Care) in Regenerative Medicine

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1097 ◽  
Author(s):  
Eckhard U. Alt ◽  
Glenn Winnier ◽  
Alexander Haenel ◽  
Ralf Rothoerl ◽  
Oender Solakoglu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Regenerative Medicine ◽  
Tissue Regeneration ◽  
Adult Stem Cells ◽  
Point Of Care ◽  
Initial Mixture ◽  
Physiological Capacity ◽  
Regenerative Cells ◽  
Regenerate Tissue

It has become practically impossible to survey the literature on cells derived from adipose tissue for regenerative medicine. The aim of this paper is to provide a comprehensive and translational understanding of the potential of UA-ADRCs (uncultured, unmodified, fresh, autologous adipose derived regenerative cells isolated at the point of care) and its application in regenerative medicine. We provide profound basic and clinical evidence demonstrating that tissue regeneration with UA-ADRCs is safe and effective. ADRCs are neither ‘fat stem cells’ nor could they exclusively be isolated from adipose tissue. ADRCs contain the same adult stem cells ubiquitously present in the walls of blood vessels that are able to differentiate into cells of all three germ layers. Of note, the specific isolation procedure used has a significant impact on the number and viability of cells and hence on safety and efficacy of UA-ADRCs. Furthermore, there is no need to specifically isolate and separate stem cells from the initial mixture of progenitor and stem cells found in ADRCs. Most importantly, UA-ADRCs have the physiological capacity to adequately regenerate tissue without need for more than minimally manipulating, stimulating and/or (genetically) reprogramming the cells for a broad range of clinical applications. Tissue regeneration with UA-ADRCs fulfills the criteria of homologous use as defined by the regulatory authorities.

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