Effects of donor age on human adipose-derived adherent stromal cells under oxidative stress conditions

Kim, Sang Woo,Choi, Jung-Won,Lee, Chang Youn,Lee, Jiyun,Shin, Sunhye,Lim, Soyeon,Lee, Seahyoung,Kim, Il-Kwon,Lee, Hoon-Bum,Hwang, Ki-Chul Cambridge Medical Publications Ltd 2018 The Journal of international medical research Vol.46 No.3

<P><B>Objective</B></P><P>Adipose-derived stromal vascular fractions (SVFs) are heterogeneous complex populations of cells with therapeutic efficacy for tissue generation and vascular stabilization. SVFs have cardiomyogenic potential, and many researchers have examined the possibility of SVF transplantation for heart disease. In cell-based therapies, donor age affects the regenerative capability, cell yield, and differentiation potential of adult tissues; however, opposing or controversial results have been found in humans. We examined whether SVF transplantation into impaired heart tissue shows differential effects according to donor age.</P><P><B>Methods</B></P><P>We investigated differences in protein expression in human umbilical vein endothelial cells (HUVECs) co-cultured with adipose-derived adherent stromal cells (ADASs) from donors of different ages [>40-year-olds (40s group) and >60-year-olds (60s group)] under oxidative stress conditions.</P><P><B>Results</B></P><P>Although co-culturing HUVECs with ADASs ameliorated inflammation due to increased oxidative stress conditions, few differences were observed between the ADASs from the 40s and 60s groups. Moreover, the Database for Annotation, Visualization, and Integrated Discovery classification tool revealed differentially expressed genes in the Kyoto Encyclopedia of Genes and Genomes pathway associated with cytokine–cytokine receptor interaction in response to ADASs.</P><P><B>Conclusion</B></P><P>Protein expression profiles were unchanged in HUVECs induced by isolated ADASs from donors of different ages under oxidative stress conditions.</P>

Rapid Induction of Osteogenic Markers in Mesenchymal Stem Cells by Adipose-Derived Stromal Vascular Fraction Cells

Choi, Jung-Won,Shin, Sunhye,Lee, Chang Youn,Lee, Jiyun,Seo, Hyang-Hee,Lim, Soyeon,Lee, Seahyoung,Kim, Il-Kwon,Lee, Hoon-Bum,Kim, Sang Woo,Hwang, Ki-Chul S. KARGER AG 2017 CELLULAR PHYSIOLOGY AND BIOCHEMISTRY Vol.44 No.1

<P>Background/Aims: Stromal vascular fraction (SVF) cells are a mixed cell population, and their regenerative capacity has been validated in various therapeutic models. The purpose of this study was to investigate the regenerative mechanisms utilized by implanted SVF cells. Using an in vitro co-culture system, we sought to determine whether SVF implantation into impaired tissue affects endogenous mesenchymal stem cell (MSC) differentiation; MSCs can differentiate into a variety of cell types, and they have a strong regenerative capacity despite their low numbers in impaired tissue. Methods: Adipose-derived SVF cells obtained from four donors were co-cultured with bone marrow-derived MSCs, and the differential expression of osteogenic markers and osteogenic differentiation inducers over time was analyzed in mono-cultured MSCs and MSCs co-cultured with SVF cells. Results: The co-cultivation of MSCs with SVF cells significantly and mutually induced the expression of osteogenic-specific markers via paracrine and/or autocrine regulation but did not induce adipocyte, chondrocyte or myoblast marker expression. More surprisingly, subsequent osteogenesis and/or comparable effects were rapidly induced within 48 h. Conclusion: To the best of our knowledge, this is the first study in which osteogenesis and/or comparable effects were rapidly induced in bone marrow-derived MSCs and adipose-derived SVF cells through co-cultivation. Our findings suggest that the positive effects of SVF implantation into impaired bone may be attributed to the rapid induction of MSC osteogenesis, and the transplantation of co-cultured and preconditioned SVF cells and/or MSCs may be more effective than the transplantation of untreated cells for the treatment of bone defects. (c) 2017 The Author(s) Published by S. Karger AG, Basel</P>

Cartilage Regeneration in Humans with Adipose Tissue-Derived Stem Cells and Adipose Stromal Vascular Fraction Cells: Updated Status

Pak, Jaewoo,Lee, Jung Hun,Pak, Natalie,Pak, Yoon,Park, Kwang Seung,Jeon, Jeong Ho,Jeong, Byeong Chul,Lee, Sang Hee MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.7

<P>Adipose tissue-derived stem cells (ASCs) in the form of stromal vascular fraction (SVF) and cultured expansion have been applied in clinical settings in some countries to treat osteoarthritis (OA) of knees, one of the most common debilitating, incurable disorders. Since the first report of successful cartilage-like tissue regeneration with autologous adipose SVF containing ASCs, there has been a gradual increase in the number of publications confirming such results. Thus far, most of the reports have been limited to treatments of OA of knees. Recently, successful applications of adipose SVF in treating OA of ankles and hips have been reported. In addition, several groups have reported modified methods of applying adipose SVF, such as combining bone marrow stimulation with adipose SVF or adding additional extracellular matrix (ECM) in treating OA. Here, we present an updated, systematic review of clinical effectiveness and safety in treating OA of knees, ankles, and one hip since 2016 using ASCs in the form of adipose SVF or in cultured expansion, along with a description and suggestion of potential biological mechanisms of cartilage regeneration.</P>

지방세포에서 비중을 이용한 지방유래 줄기세포가 풍부한 분획의 분리 방법

김민경,박용순,박희순,최정묵,김원준,박세은,이은정,박철영,이원영,오기원,박성우,김선우,서광식,우정택 대한내분비학회 2010 Endocrinology and metabolism Vol.25 No.2

Background: Adipose tissues include multipotent cells, the same as bone marrow-derived mesenchymal stem cells. The stromal vascular fractions (SVFs) from adipose tissues represent a heterogeneous cell population. The purpose of this study was to isolate and purify adipose-derived stem cells (ASCs) in SVFs by the density gradient method. Methods: SVFs were extracted from the subcutaneous, epididymal, mesenteric and retroperitoneal adipose tissue of 8 weeks old male Sprague-Dawley rats (n = 15) and these were separated into 4 layers according to a Nycodenz gradient (Fx-1: < 11%, Fx-2: 11-13%, Fx-3: 13-19% and Fx-4: 19-30%). The post-confluent SVFs were cultured in adipogenic medium for 2 days, in insulin medium for 2 days and in 10% fetal bovine serum medium for 5 days. To observe lipid droplets in SVFs, we performed Oil Red O staining. Results: The SVFs’ cellular fractions (Fx-1, Fx-2, Fx-3 and Fx-4) were isolated by density gradient centrifugation from the adipose tissues of rats. The SVFs extracted to fraction 3 (Fx-3) had the most abundant cells compared to that of the other fractions. However fraction 1 (Fx-1) or 2 (Fx-2) had a superior ability to make lipid droplets. The adipogenic differentiation of Fx-1 or 2 was higher than that of the unfractionated cells. The SVFs extracted from retroperitoneal adipose tissue had the highest efficiency for adipogenic differentiation,whereas the SVFs from mesenteric adipose tissue did not differentiate. Conclusion: This density gradient fractionated method leads to efficient isolation and purification of cells with the characteristics of ASCs. 배경: 지방조직은 골수 유래의 중간엽 줄기세포와 같은 다분화 능력을 가진 성체줄기세포를 포함한다. 지방조직으로부터 추출한 지방유래 줄기세포가 다양한 분화 유도를 통해 지방세포, 골세포, 근육세포로 분화할 수 있음이 밝혀졌다. 본 연구에서는 지방조직으로부터 얻은 기질세포에서 지방세포로의 탁월한 분화능력을 가지고 있는 순수한 지방유래 줄기세포를 추출해 낼 수 있는 지 알아보았다. 방법: 수컷 Sprague-Dawley 쥐(n = 15)의 4부위 지방조직(피하 지방, 부고환 지방, 장간막 지방, 후복막 지방)에서 불필요한 조직을 제거한 나머지 기질세포만을 얻는다. Nycodenz를 이용하여 기질세포를 밀도차등원심분리하여 4개의 분획을 얻어 각각을 배양시킨다.포화 상태에 이른 지방줄기세포를 지방세포로 분화를 유도하여 세포 내 지방 축적 여부를 염색을 통해 확인한다. 결과: 밀도차등 원심분리로 지방조직 기질세포는 4개의 분획(Fx-1, Fx-2, Fx-3, Fx-4)으로 나뉘었으며, 4부위 지방조직 모두 세 번째분획(Fx-3)의 세포 수가 가장 많았다. 그러나 지방세포로의 분화는 4부위 지방조직 모두 두 번째 분획(Fx-2)에서 탁월했으며 분획을 나누지 않은 것(control)과 비교했을때 더 나은 분화 능력을 보였다.지방으로 분화가 가장 잘 되는 지방 조직은 retroperitoneal fat tissue였으며 그 다음이 subcutaneous fat tissue였고, epididymal fat tissue는 상대적으로 분화율이 떨어지는 것을 관찰하였다. Mesenteric fattissue는 분화가 거의 되지 않았다. 결론: 지방조직별, 밀도별로 지방조직 기질세포를 나누어 분리함으로써 지방조직으로의 분화 능력이 우수한 지방줄기세포를 찾을수 있었다. 향후 지방줄기세포를 이용한 연구 및 활용에 있어서 지방조직별, 분획별 기질세포의 이용으로 일관된 결과를 얻는 데 도움이 될 것이다.

Stem cells for cartilage repair: what exactly were used for treatment, cultured adipose-derived stem cells or the unexpanded stromal vascular fraction?

Lee, J.I.,Balolong, E.,Han, Y.,Lee, S. Published for the Society by Baillère Tinda 2016 OSTEOARTHRITIS AND CARTILAGE Vol.24 No.7

Reconsidering cell-assisted lipotransfer for breast augmentation: effect of stromal vascular fraction enrichment on graft survival assessed with 3-dimensional laser scanning

Li-Chen Chiu,Li-Shu Chiu,Cheng-Hung Chiu 대한미용의학회 2019 대한미용의학회지 Vol.3 No.1

Background: Cell-assisted lipotransfer (CAL) has been proposed to be beneficial for improving graft retention. Clinically, CAL involves the isolation of the stromal vascular fraction (SVF) from a portion of the lipoaspirate at the time of surgery. However, most studies related to SVF breast augmentation lacked a rigorous methodology and well-designed control. Objective: We aimed to determine the potential improvement of SVF enrichment in fat grafting for breast augmentation with objective volume assessment. Methods: From April 2015 to January 2016, 169 patients were enrolled after applying the exclusion criteria. Among them, 97 patients who underwent conventional fat grafting for breast augmentation were assigned to group A. The other 72 patients underwent SVFenriched fat grafting for breast augmentation and were assigned to group B. A retrospective comparative study was conducted to evaluate the graft survival using 3-dimensional laser scanning. Results: There was no significant difference between the 2 groups in terms of mean age, original breast volume, grafted fat volume, and postoperative weight change. Breast volume assessments revealed that the percentage of graft survival at 12 months was 69.2% in group A and 71.1% in group B, with no significant difference (p=0.641). The preoperative body mass index was significantly lower in group A than in group B. The volume of suctioned fat was significantly less in group A. The operation time was significantly shorter in group A. The postoperative complication rates were significantly lower in group A than in group B. Conclusion: SVF-enriched fat grafting for breast augmentation was associated with a larger amount of harvested fat, a longer operation time, and a higher incidence of complications. The graft retention rate was not significantly increased. The findings of our study do not support the use of SVF in fat grafting for breast augmentation. Level of Evidence: IV

Effect of Collagen Scaffold with Adipose-derived Stromal Vascular Fraction Cells on Diabetic Wound Healing: A Study in a Diabetic Porcine Model

Yanan Jiang,Bing Chen,Yongbo Liu,Zhongyin Zhufu,Xin Yan,Xianglin Hou,Jianwu Dai,Qian Tan 한국조직공학과 재생의학회 2013 조직공학과 재생의학 Vol.10 No.4

Diabetes can impair neovascularization important for wound healing, and this study investigated the hypothesis that this effect can be reversed by an active provisional matrix. Here, a 3D collagen scaffold and uncultured adipose-derived SVFs were firstly utilized to construct a collagen-targeting system for wound healing in a diabetic porcine model. The diabetic porcine models were made by injected streptozocin (STZ) intravenously and removed full thickness skin from the dorsum. Each animal received four different samples: Group A (control groups, n=24), Group B (SVFs groups, n=24), Group C (scaffold groups, n=24), Group D (SVFs -scaffold groups, n=24). PBS was applied to the wounds in Group A. 1×105 SVFs suspended in PBS were sprayed on the wounds in Group B. Scaffolds only with PBS were covered on the wounds in Group C. Scaffolds with equal volume of cell suspension containing 1×105 SVFs were covered on the wounds in Group D. The healing rates were calculated and compared among the groups and the tissues of the wound were taken and evaluated for histological analysis. The diabetic wound treated with the SVFs -collagen scaffold showed a significant decrease in wound size, an increase of neovascularization and a maximum VEGF and bFGF expression in comparison with SVFs and scaffold alone. The SVFs-collagen scaffold accelerates wound healing since it stimulates higher capillary formation causes major proangiogenesis factors expression in diabetic pigs suffering from skin defects, suggesting this approach could have utility for diabetic wound healing.

Current use of autologous adipose tissue-derived stromal vascular fraction cells for orthopedic applications

Pak, Jaewoo,Lee, Jung Hun,Park, Kwang Seung,Park, Moonhee,Kang, Lin-Woo,Lee, Sang Hee BioMed Central 2017 JOURNAL OF BIOMEDICAL SCIENCE -BASEL- Vol.24 No.1

<P>Autologous adipose stromal vascular fractions (SVFs) containing adipose tissue-derived stem cells (ASCs) are currently being used in clinical settings for various orthopedic applications for human patients. Due to its potential capability of regenerating cartilage, bone, and tendons, autologous adipose SVFs are being tried in treating patients with osteoarthritis (OA), chondromalacia, meniscus tear, osteonecrosis of the femoral head, and tendon injuries. Here, we have reviewed available human clinical studies with regard to patient applications of autologous adipose SVF containing ASCs, specifically assessing effectiveness and safety in the field of orthopedic disorders. All studies reviewed in this article presents potential benefits of autologous adipose SVF in various orthopedic applications without any serious side effects.</P>

Complex sternal and rib reconstruction with allogeneic material

Charles Miles Maliska III,Robert Lloyd Archer,Sharon Kaye Tarpley 대한성형외과학회 2018 Archives of Plastic Surgery Vol.45 No.6

Sternal malunion, or loss, developed after a median sternotomy cannot only be difficult to manage and treat, but also may diminish one’s quality-of-life drastically. The technique presented here represents a multispecialty approach in one stage for the reconstruction of an unstable thoracic cage. The procedure utilized a donated sternum and ribs. The sternum with ribs harvested from a single donor included adipose derived stromal vascular fraction (ADSVF) cells with marrow also from the same donor. Autologous muscle flaps, stabilized with acellular dermal matrix were utilized to provide a robust blood supply to the ADSVF cells and bone grafts. Acellular dermal matrix was used to construct the ribs and stabilize the plugs of stem cells and bone. These procedures, in the hands of multispecialty physicians, have led to several successful reconstructions involving complex chest wall deformities. This surgical intervention was performed in a one stage operation. This represents the first successful complete sternal transplant in a patient with return to normal activities and increased quality-of-life.

Complex sternal and rib reconstruction with allogeneic material

Maliska, Charles Miles III,Archer, Robert Lloyd,Tarpley, Sharon Kaye,Miller III, Archibald Sanford Korean Society of Plastic and Reconstructive Surge 2018 Archives of Plastic Surgery Vol.45 No.6

Sternal malunion, or loss, developed after a median sternotomy cannot only be difficult to manage and treat, but also may diminish one's quality-of-life drastically. The technique presented here represents a multispecialty approach in one stage for the reconstruction of an unstable thoracic cage. The procedure utilized a donated sternum and ribs. The sternum with ribs harvested from a single donor included adipose derived stromal vascular fraction (ADSVF) cells with marrow also from the same donor. Autologous muscle flaps, stabilized with acellular dermal matrix were utilized to provide a robust blood supply to the ADSVF cells and bone grafts. Acellular dermal matrix was used to construct the ribs and stabilize the plugs of stem cells and bone. These procedures, in the hands of multispecialty physicians, have led to several successful reconstructions involving complex chest wall deformities. This surgical intervention was performed in a one stage operation. This represents the first successful complete sternal transplant in a patient with return to normal activities and increased quality-of-life.