Human adipose derived mesenchymal stem cell

Human adipose-derived mesenchymal stem cells (hASCs) have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation.

hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time.

The potential therapeutic benefit of adipose-derived stem cells (ASCs) encapsulated in an injectable hydrogel for stimulating intervertebral disc (IVD) regeneration has been assessed by a number of translational and preclinical studies. However, previous work has been primarily limited to small animal models and short-term outcomes of only a few weeks. Long-term studies in representative large animal models are crucial for translation into clinical success, especially for permanent stabilization of major defects such as disc herniation. An injectable chitosan carboxymethyl cellulose hydrogel scaffold loaded with ASCs was evaluated regarding its intraoperative handling, crosslinking kinetics, cell viability, fully-crosslinked viscoelasticity, and long-term therapeutic effects in an ovine model. Three IVDs per animal were damaged in 10 sheep. Subcutaneous adipose tissue was the source for autologous ASCs. Six weeks after IVD damage, two of the damaged IVDs were treated via ASC-loaded hydrogel injection. After 12 months following the implantation, IVD disc height and histological and cellular changes were assessed. This system was reliable and easy to handle intraoperatively. Over 12 months, IVD height was stabilized and degeneration progression significantly mitigated compared to untreated, damaged IVDs. Here we show for the first time in a large animal model that an injectable chitosan carboxymethyl cellulose hydrogel system with encapsulated ASCs is able to affect long-term stabilization of an injured IVD and significantly decrease degeneration processes as compared to controls 1).

Chen et al. performed intracerebroventricular microinjection of human adipose mesenchymal stem cell (hADSC)-derived exosomes (hADSC-ex) in a weight-drop-induced TBI rat model.

They found that hADSC-ex promoted functional recovery, suppressed neuroinflammation, reduced neuronal apoptosis, and increased neurogenesis in TBI rats. The therapeutic effects of hADSC-ex were comparable to those of hADSC. Sequential in vivo imaging revealed increasing aggregation of DiR-labeled hADSC-ex in the lesion area. Immunofluorescent staining of coronal brain sections and primary mixed neural cell cultures revealed distinct overlap between CM-DiI-labeled hADSC-ex and microglia/macrophages, indicating that hADSC-ex were mainly taken up by microglia/macrophages. In a lipopolysaccharide-induced inflammatory model, hADSC-ex suppressed microglia/macrophage activation by inhibiting nuclear factor κB and P38 mitogen-activated protein kinase signaling. These data suggest that hADSC-ex specifically enter microglia/macrophages and suppress their activation during brain injury, thereby inhibiting inflammation and facilitating functional recovery. They also offer new insight into the cellular targeting, uptake and migration of hADSC-ex, and provide a theoretical basis for new therapeutic strategies for central nervous system diseases 2).

Three patients (two patients and one patient with 1 and 2 levels, respectively) with degenerative spondylolisthesis underwent MI-TLIF with 3D graft made of AMSCs. To obtain the AMSCs, fatty tissue was collected from the abdomen by lipoaspiration and differentiated afterwards in our Cell/Tissue bank. Clinical outcomes, including the Oswestry Disability Index (ODI) and visual analog scale (VAS) as well as fusion status were assessed preoperatively and up to 12 months postoperatively.

At 12 months, all four operated AMSC levels could be assessed (n = 4). Grade 3 fusion could be confirmed at two levels out of four. Mean VAS score improved from 8.3 to 2 and ODI also improved from 47 to 31%. No donor site complication was observed. The final AMSC osteogenic product was stable, did not rupture with forceps manipulation, and was easily implanted directly into the cage with no marked modification of operating time.

A scaffold-free 3D graft made of AMSCs can be manufactured and used as a promising alternative for spinal fusion procedures. Nevertheless, further studies of a larger series of patients are needed to confirm its effectiveness. 3).

Matrix metalloproteinases (MMPs) are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells.

Matrix metalloproteinases (MMPs) secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs 4).

El Bassit et al., evaluated the effect of hASC secretome on HT22 neuronal cells post injury. Protein Kinase C delta (PKCδ)1 activates survival and proliferation in neurons and is implicated in memory (1-3). We previously showed that alternatively spliced PKCδII enhances neuronal survival via Bcl2 (4) in HT22 neuronal cells. Our results demonstrate that following injury, treatment with exosomes from the hASC secretome increases expression of PKCδII in HT22 cells and increases neuronal survival and proliferation. Specifically, we demonstrate that MALAT1, a long noncoding RNA contained in the hASC exosomes mediates PKCδII splicing thereby increasing neuronal survival. Using antisense oligonucleotides for MALAT1 and RNA immunoprecipitation assays we demonstrate that MALAT1 recruits splice factor SRSF2 to promote alternative splicing of PKCδII. Finally, we evaluated the role of insulin in enhancing hASC mediated neuronal survival, and demonstrated that insulin treatment dramatically increased the association of MALAT1 and SRSF2 and substantially increases survival and proliferation after injury in HT22 cells. In conclusion, we demonstrate the mechanism of action of hASC exosomes in increasing neuronal survival. This effect of hASC exosomes to promote wound healing can be further enhanced by insulin treatment in HT22 cells 5).

human adipose tissue-derived mesenchymal stem cells (ADSCs) secrete exosomes carrying enzymatically active NEP. The NEP-specific activity level of 1 μg protein from ADSC-derived exosomes was equivalent to that of ~ 0.3 ng of recombinant human NEP. Of note, ADSC-derived exosomes were transferred into N2a cells, and were suggested to decrease both secreted and intracellular Aβ levels in the N2a cells. Importantly, these characteristics were more pronounced in ADSCs than bone marrow-derived mesenchymal stem cells, suggesting the therapeutic relevance of ADSC-derived exosomes for AD 6).

Schmitt C, Radetzki F, Stirnweiss A, Mendel T, Ludtka C, Friedmann A, Baerthel A, Brehm W, Milosevic J, Meisel HJ, Goehre F, Schwan S. Long-term Pre-Clinical Evaluation of an Injectable Chitosan Nanocellulose Hydrogel with Encapsulated Adipose-Derived Stem Cells in an Ovine Model for IVD Regeneration. J Tissue Eng Regen Med. 2021 May 14. doi: 10.1002/term.3216. Epub ahead of print. PMID: 33989456.
Chen Y, Li J, Ma B, Li N, Wang S, Sun Z, Xue C, Han Q, Wei J, Zhao RC. MSC-derived exosomes promote recovery from traumatic brain injury via microglia/macrophages in rat. Aging (Albany NY). 2020 Sep 23;12. doi: 10.18632/aging.103692. Epub ahead of print. PMID: 32966240.
Fomekong E, Dufrane D, Berg BV, André W, Aouassar N, Veriter S, Raftopoulos C. Application of a three-dimensional graft of autologous osteodifferentiated adipose stem cells in patients undergoing minimally invasive transforaminal lumbar interbody fusion: clinical proof of concept. Acta Neurochir (Wien). 2016 Dec 30. doi: 10.1007/s00701-016-3051-6. [Epub ahead of print] PubMed PMID: 28039550.
Arai Y, Park S, Choi B, Ko KW, Choi WC, Lee JM, Han DW, Park HK, Han I, Lee JH, Lee SH. Enhancement of Matrix Metalloproteinase-2 (MMP-2) as a Potential Chondrogenic Marker during Chondrogenic Differentiation of Human Adipose-Derived Stem Cells. Int J Mol Sci. 2016 Jun 17;17(6). pii: E963. PubMed PMID: 27322256.
El Bassit G, Patel RS, Carter G, Shibu V, Patel A, Song S, Murr M, Cooper DR, Bickford PC, Patel NA. MALAT1 in human adipose stem cells modulates survival and alternative splicing of PKCδII in HT22 cells. Endocrinology. 2016 Nov 14:en20161819. [Epub ahead of print] PubMed PMID: 27841943.
Katsuda T, Tsuchiya R, Kosaka N, Yoshioka Y, Takagaki K, Oki K, Takeshita F, Sakai Y, Kuroda M, Ochiya T. Human adipose tissue-derived mesenchymal stem cells secrete functional neprilysin-bound exosomes. Sci Rep. 2013;3:1197. doi: 10.1038/srep01197. Epub 2013 Feb 1. PubMed PMID: 23378928; PubMed Central PMCID: PMC3561625.
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