Human Stem Cell Therapy Research Articles

Chemically Defined Media Can Maintain Pig Pluripotency Network InVitro.

SummaryPig embryonic stem cells (pESCs) have been considered an important candidate for preclinical research on human therapies. However, the lack of understanding of pig pluripotent networks has hampered establishment of authentic pESCs. Here, we report that FGF2, ACTVIN, and WNT signaling are essential to sustain pig pluripotency in vitro. Newly derived pESCs were stably maintained over an extended period, and capable of forming teratomas that contained three germ layers. Transcriptome analysis showed that pESCs were developmentally similar to late epiblasts of preimplantation embryos and in terms of biological functions resembled human rather than mouse pluripotent stem cells. However, the pESCs had distinct features such as coexpression of SSEA1 and SSEA4, two active X chromosomes, and a unique transcriptional pattern. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use.

PPIA, HPRT1, and YWHAZ Genes Are Suitable for Normalization of mRNA Expression in Long-Term Expanded Human Mesenchymal Stem Cells.

Long-term expansion of mesenchymal stem cells (MSCs) under defined culture conditions is necessary in human stem cell therapy. However, it alters the characteristics of MSCs. Since quantitative real time polymerase chain reaction (qRT-PCR) is widely used as one of the key analytical methods for comparative characterization, the validation of reference genes (RGs) for normalization under each experimental condition is important to achieve reliable qRT-PCR results. Therefore, the most stable RGs for long-term expanded bone marrow- and umbilical cord blood-derived MSCs (BM-MSCs and UCB-MSCs) under defined culture conditions for up to 20 passages were evaluated. The more apparent alterations in characteristics such as differentiation capacity, proliferation, senescence, and the expression of RGs were noted in BM-MSCs than UCB-MSCs during long-term expansion. The RG validation programs (GeNorm and NormFinder) suggested that PPIA, HPRT1, and YWHAZ were suitable for normalization in qRT-PCR regardless of MSC types and long-term culture expansion, and the traditional RGs (ACTB and GAPDH) were less stable in long-term expanded MSCs. In addition, the use of these RGs for normalization of OCT4 expression in long-term expanded BM-MSCs showed that a less stable RG (GAPDH) showed contrasting data compared to other RGs. Therefore, the use of RGs such as PPIA, HPRT1, and YWHAZ for normalization in qRT-PCR experiments is highly recommended for long-term expanded MSCs to generate accurate and reliable data.

Developing a human mesenchymal stem cell culture medium with high growth efficient, 3d culture supported, serum-free and xeno-free properties.

Background & Aim Human mesenchymal stem cell (MSC) therapies have been tested clinically for a variety of disorders, including Crohn's disease, multiple sclerosis, graft-versus-host disease (GVHD), bone fractures, sepsis, and cartilage defects. Despite of the remarkable clinical advancements in this field, most cell manufacturing processes still rely on the traditional culture medium containing fetal bovine serum (FBS). The highly inconsistence nature and xenogenic infection risks of the bovine serum remain a challenge for the standardization and massive production for clinics. Here, we successfully develop a medium, featured with serum-free, xeno-free components and high efficacy for MSC growth, named as AllPhase. Methods, Results & Conclusion The medium on human adipose tissue-derived mesenchymal stem cells (ADSC) and Wharton's jelly mesenchymal stem cells (WJ-MSC) enables competitive expansion performance, comparing to traditional culture medium and can maintains tri-lineage mesoderm differentiation potential after fifth passages. Furthermore, most commercial serum-free media need expansive ECM coating, like fibronectin or other commercial coating materials, to support cell adhesion. Especially, AllPhase can support ADSC expansion under 3D culture condition without microcarriers. We hope that our medium can support most bioreactors and cell industrial production for MSC clinical applications.

Human Placental Stem Cell Therapy in Stroke: Endothelial / Smooth Muscle Mechanisms Underlying Protection?

Following thrombolytic therapy for stroke, ischemia/reperfusion (I/R) mediated inflammation often disrupts the blood brain barrier (BBB). This can enhance expression of endothelial adhesion markers and perturb normal blood flow regulation. Proposed benefits of stem cell therapy (SCT) in stroke include secretion of protective and trophic factors which limit acute brain I/R injury. We evaluated human placenta mesenchymal stem cells (hPMSC) as potential ameliorative SCT in an acute ischemic stroke model. We hypothesized that potential benefits of hPMSCs, in acute stroke injury, reflect site‐specific suppression of post‐ischemic immune cell adhesion, transvascular leukocyte emigration and preservation of the BBB. This can limit the progression of post –ischemic stroke injury. We found that intraperitoneal (IP) administration of hPMSCs at the time of reperfusion, using the transient middle cerebral artery occlusion (MCAO)/reperfusion model, produced highly significant protection of the ipsilateral hemisphere. We then examined hPMSC support of BBB integrity via mobilization of junctional proteins in human brain endothelial cells (hCMEC‐D3) in conjunction with a biotin/FITC‐avidin permeability assay coupled with western blotting and immunolocalization of junctional proteins. We also assessed the ability of hPMSC to suppress inflammation‐driven hCMEC‐D3 binding of macrophages (RAW264.7) and transmigration of PMNs. In addition, we used collagen gels to study the influence of hPMSC on human brain vascular smooth muscle cells (hBVSMC) tonic contractility.We observed that hPMSC administration significantly (p<0.001) enhanced the barrier integrity of brain endothelial monolayers, at 24h, in an oxygen/glucose deprived/reperfused (OGDR) condition. We found that hCMEC‐D3, which were independently co‐cultured with hPMSCs, expressed significantly higher levels of VE‐cadherin and α‐catenin under OGDR conditions (p<0.001). Additionally, the spatial localization of VE‐cadherin and α‐catenin at hCMEC‐D3 junctions was increased, in the OGDR state, following hPMSC treatment. We also observed that hPMSC treatment significantly (p<0.0001) reduced binding of RAW264.7 to TNF‐α/ LPS activated hCMEC‐D3 monolayers. The expression of VCAM‐1 was increased in TNF‐α/ LPS activated hCMEC‐D3 while this was significantly (p<0.01, P<0.05) decreased by hPMSC treatment. hPMSC also inhibited (p<0.05) PMN transmigration across hCMEC‐D3 monolayers. Lastly, we found that hBVSMC contraction was significantly increased by hPMSC treatment (p=0.0001). This effect was reversed by angiotensin II Type 1 receptor blockade (Losartan, 50μM), angiotensin converting enzyme (Captopril, 50μM), thromboxane A2 synthesis (Ozagrel, 10μM), and endothelin converting enzyme inhibition (SM19712, 100μM).We conclude that pleiotropic factors associated with hPMSC administration can have a favorable impact on BBB integrity, immune cell interaction, and vasoregulation potentially alleviating the detrimental effects of ischemic stroke.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 Diabetes and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture

BackgroundPrevious work showed that muscle-derived stem cells (MDSCs) exposed long-term to the milieu of uncontrolled type 2 diabetes (UC-T2D) in male obese Zucker (OZ) rats, were unable to correct the associated erectile dysfunction and the underlying histopathology when implanted into the corpora cavernosa, and were also imprinted with a noxious gene global transcriptional signature (gene-GTS), suggesting that this may interfere with their use as autografts in stem cell therapy. AimTo ascertain the respective contributions of dyslipidemia and hyperglycemia to this MDSC damage, clarify its mechanism, and design a bioassay to identify the damaged stem cells. Methods: Early diabetes MDSCs and late diabetes MDSCs were respectively isolated from nearly normal young OZ rats and moderately hyperglycemic and severely dyslipidemic/obese aged rats with erectile dysfunction. Monolayer cultures of early diabetic MDSCs were incubated 4 days in DMEM/10% fetal calf serum + or − aged OZ or lean Zucker serum from non-diabetic lean Zucker rats (0.5–5%) or with soluble palmitic acid (PA) (0.5–2 mM), cholesterol (CHOL) (50–400 mg/dL), or glucose (10–25 mM). Main Outcome MeasureFat infiltration was estimated by Oil red O, apoptosis by TUNEL, protein expression by Western blots, and gene-GTS and microRNA (miR)-GTS were determined in these stem cells’ RNA. ResultsAged OZ serum caused fat infiltration, apoptosis, myostatin overexpression, and impaired differentiation. Some of these changes, and also a proliferation decrease occurred with PA and CHOL. The gene-GTS changes by OZ serum did not resemble the in vivo changes, but some occurred with PA and CHOL. The miR-GTS changes by OZ serum, PA, and CHOL resembled most of the in vivo changes. Hyperglycemia did not replicate most alterations. Clinical Implications: MDSCs may be damaged in long-term UC-T2D/obese patients and be ineffective in autologous human stem cell therapy, which may be prevented by excluding the damaged MDSCs. Strength & LimitationsThe in vitro test of MDSCs is innovative and fast to define dyslipidemic factors inducing stem cell damage, its mechanism, prevention, and counteraction. Confirmation is required in other T2D/obesity rat models and stem cells (including human), as well as miR-GTS biomarker validation as a stem cell damage biomarker. ConclusionSerum from long-term UC-T2D/obese rats or dyslipidemic factors induces a noxious phenotype and miR-GTS on normal MDSCs, which may lead in vivo to the repair inefficacy of late diabetic MDSCs. This suggests that autograft therapy with MDSCs in long-term UT-T2D obese patients may be ineffective, albeit this may be predictable by prior stem cell miR-GTS tests.Masouminia M, Gelfand R, Kovanecz I, et al. Dyslipidemia Is a Major Factor in Stem Cell Damage Induced by Uncontrolled Long-Term Type 2 Diabetes and Obesity in the Rat, as Suggested by the Effects on Stem Cell Culture. J Sex Med 2018;15:1678–1697.

Abstract 1966: A novel small-molecule compound targeting PBX1-DNA interaction impedes cancer cell survival and carboplatin resistance

Abstract Pre-B-cell leukemia homeobox-1 (PBX1), a transcriptional factor and downstream effector of Notch signaling pathway, plays pivotal roles in a wide spectrum of tumors, as well as developmental, inflammatory, autoimmune, and neurodegenerative disorders. Exploiting the crystal structure of the Pbx1-DNA complex, we developed a novel small-molecular inhibitor T417 that can directly block Pbx1-binding to DNA, unlike other existing compounds that interfere with protein-protein interactions. When T417 docks into the hydrophobic pocket of Pbx1 protein, the small molecule can dampen PBX1 transcription activity by hindering its binding to the promoter regions of PBX1 downstream target genes. Intriguingly, the amount of PBX1 expression in cells can dictate its response to T417. Increment of PBX1 expression is found in the high-grade serous carcinoma (HGSC) and carboplatin-resistant (CR) cells, and its expression is generally low in normal tissues as compared to the transformed tissues, making it such an arguably unique therapeutic target in ovarian cancer cells. This expression pattern illustrates the very minimal toxicity of T417 on normal tissues and organs in animal models, while it imposed in vitro and in vivo detrimental effects toward HGSC and CR cells. Besides, T417 holds synergistic cytotoxic effects with DNA damage-related drugs including PARP inhibitor and platinum-based drug. As PBX1 was shown to participate in maintaining cancer stem cell (CSC)-like phenotypes and promoting resistance to antitumor drugs, T417 is able to hammer out the stemness traits of CR cells to revert to a differentiated status through tacking PBX1 signaling cascade. The novel PBX1-targeting compound selectively interferes with PBX1-binding to DNA, which potentially points to powerful therapeutics and broad applications for the treatment of different human malignancies and stem cell therapy. Citation Format: Yao-An Shen, Jin Jung, Yohan Suryo Rahmanto, Licia Selleri, Ie-Ming Shih, Chi-Mu Chuang, Tian-Li Wang. A novel small-molecule compound targeting PBX1-DNA interaction impedes cancer cell survival and carboplatin resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1966.

Advantages of Graphene Biosensors for Human Stem Cell Therapy Potency Assays.

Regenerative medicine is challenged by the need to conform to rigorous guidelines for establishing safe and effective development and translation of stem cell-based therapies. Counteracting widespread concerns regarding unproven cell therapies, stringent cell-based assays seek not only to avoid harm but also to enhance quality and efficacy. Potency indicates that the cells are functionally fit for purpose before they are administered to the patient. It is a paramount quantitative critical quality attribute serving as a decisive release criterion. Given a broad range of stem cell types and therapeutic contexts the potency assay often comprises one of the most demanding hurdles for release of a cell therapy medicinal product. With need for improved biomarker assessment and expedited measurement, recent advances in graphene-based biosensors suggest that they are poised to be valuable platforms for accelerating potency assay development. Among several potential advantages, they offer versatility for sensitive measurement of a broad range of potential biomarker types, cell biocompatibility for direct measurement, and small sample sufficiency, plus ease of use and point-of-care applicability.

A working model for hypothermic neuroprotection.

Therapeutic hypothermia significantly improves survival without disability in near-term and full-term newborns with moderate to severe hypoxic-ischaemic encephalopathy. However, hypothermic neuroprotection is incomplete. The challenge now is to find ways to further improve outcomes. One major limitation to progress is that the specific mechanisms of hypothermia are only partly understood. Evidence supports the concept that therapeutic cooling suppresses multiple extracellular death signals, including intracellular pathways of apoptotic and necrotic cell death and inappropriate microglial activation. Thus, the optimal depth of induced hypothermia is that which effectively suppresses the cell death pathways after hypoxia-ischaemia, but without inhibiting recovery of the cellular environment. Thus mild hypothermia needs to be continued until the cell environment has recovered until it can actively support cell survival. This review highlights that key survival cues likely include the inter-related restoration of neuronal activity and growth factor release. This working model suggests that interventions that target overlapping mechanisms, such as anticonvulsants, are unlikely to materially augment hypothermic neuroprotection. We suggest that further improvements are most likely to be achieved with late interventions that maximise restoration of the normal cell environment after therapeutic hypothermia, such as recombinant human erythropoietin or stem cell therapy.

Correction: Human adipose derived stem cells regress fibrosis in a chronic renal fibrotic model induced by adenine

[This corrects the article DOI: 10.1371/journal.pone.0187907.].

Regulating human stem cell research and therapy in low- and middle-income countries: Malaysian perspectives

Many “rising powers” such as India, China, Argentina, Singapore, and Brazil are investing in stem cell technology, joining the traditional leaders in the field, such as the UK, Germany, USA, and Japan. Malaysia is also entering this sector because of the potential medical and economic benefits that the use of stem cell technologies could provide. Like other countries, Malaysia faces the challenge of how to encourage scientific progress and innovation in an ethical manner while at the same time ensuring a safe and accessible market for regenerative therapies. This paper reports on the research findings of semi-structured interviews with local stakeholders to investigate how they perceived and evaluated the current regulatory framework for human stem cell research in Malaysia, and what might be at stake if the state continues with its current regulatory approach.

Human adipose derived stem cells regress fibrosis in a chronic renal fibrotic model induced by adenine.

Background and aimsADSCs transplantation had been shown in some experimental models of kidney damage that it improves kidney function and reduces fibrosis. In this study we evaluated the effect of human adipose tissue-derived stem cell (hADSC) therapy in a chronic kidney damage experimental model.MethodsA chronic kidney injury was induced by daily orogastric administration of adenine (100mg/kg) to male Wistar rats for 28 days. hADSCs were isolated, expanded and characterized before transplantation. hADSC administration was performed in a tail vein at a dose of 2 x106 cells/animal. Animals were sacrificed at 7 days post-treatment. The percentage of fibrotic tissue, serum and urine levels of urea, creatinine, total protein and renal mRNA of COL1A1, TGFB1, CTGF, ACTA2, IL6, IL10, TNF were analyzed.ResultshADSCs treatment significantly reduces kidney fibrosis, improves urea and creatinine serum and urine levels, and diminishes COL1A1, TGFB1, CTGF, ACTA2 mRNA kidney levels.ConclusionsThese results showed that cell therapy using hADSCs improves renal function and reduces fibrosis.

Human Stem Cell Therapy Is Successful in Monkeys with Parkinson Disease

An individual's induced pluripotent stem (iPS) cells can be coaxed into becoming specialized cells to replace cells lost to disease (NEJM JW Gen Med

ID: 1041 Effectiveness of human adipose-derived stem cell therapy pretreated with hepatocyte growth factor in liver fibrosis mouse model

Background: Adipose-derived stem cells (ADSCs) have the potential therapeutic impact on the liver fibrosis. Hepatocyte growth factor (HGF) is pivotal for damage repair with its anti-apoptotic, anti-fibrotic and cell migration-promoting effect. In this study, the therapy with HGF-pretreated hADSCs was expected to enhance the therapeutic effect in the amelioration of liver fibrosis compared with untreated hADSCs.
 
 Method: HGF-hADSCs were prepared by culturing hADSCs for seven days in the medium added HGF.
 HGF-hADSCs transplantation was performed to investigate the therapeutic effect of these cells on carbon tetrachloride (CCl4)-induced liver fibrosis in a mouse model. 
 Results: After seven days and fourteen days of cell transfusion, HGF-hADSCs ameliorated the liver fibrosis. The results showed the attenuation of the liver injury (ALT level), the down-regulation of procollagen-1 (7 days, 3-fold; 14 days, 6.7-fold) and alpha -smooth muscle actin (alpha-SMA) expression (7 days, 10-fold; 14 days, 2-fold), and the histological improvement. Notably, there was the significant difference in the procollagen-1 between HGF-hADSCs and untreated hADSCs groups. Thus, the therapy with HGF-hADSCs was more efficient in the liver fibrosis treatment compared with untreated hADSCs. 
 Conclusion: HGF pretreated hADSCs have a potential therapy in the treatment of liver fibrosis

Human Neural Stem Cell Therapy for Chronic Ischemic Stroke: Charting Progress from Laboratory to Patients.

Chronic disability after stroke represents a major unmet neurologic need. ReNeuron's development of a human neural stem cell (hNSC) therapy for chronic disability after stroke is progressing through early clinical studies. A Phase I trial has recently been published, showing no safety concerns and some promising signs of efficacy. A single-arm Phase II multicenter trial in patients with stable upper-limb paresis has recently completed recruitment. The hNSCs administrated are from a manufactured, conditionally immortalized hNSC line (ReNeuron's CTX0E03 or CTX), generated with c-mycERTAM technology. This technology has enabled CTX to be manufactured at large scale under cGMP conditions, ensuring sufficient supply to meets the demands of research, clinical development, and, eventually, the market. CTX has key pro-angiogenic, pro-neurogenic, and immunomodulatory characteristics that are mechanistically important in functional recovery poststroke. This review covers the progress of CTX cell therapy from its laboratory origins to the clinic, concluding with a look into the late stage clinical future.

Sub-national technology policy and commerce: evaluating the impacts of the California Institute for Regenerative Medicine

In the last 20 years, state governments have funded a large number of science-based economic development programs. Remarkably, there have been few evaluations of either their economic impact or their prospects for economic impact. The 2004 decision by California voters to issue bonds to fund $3 billion of research on human pluripotent stem cells is an ideal case study for introducing an evaluation methodology for the progress by state programs in developing and transferring research results. In the process of evaluating the California stem cell initiative, we make three methodological contributions to the study of the impacts of sub-national science-based economic development programs. First, the following data sources are introduced as indicators of the prospective economic benefits of these programs: changes in targeted federal research funding, patenting, small business investment research grants, venture capital investment, new firm formations, and clinical trials. Second, the use of regional or industrial controls is introduced. In the case study of the California stem cell initiative the regional control is the comparison of the two dominant California stem cell research and firm clusters, San Diego and the San Francisco Bay Area, to the other dominant cluster, Boston. The biotechnology industry, as a whole, is used to control for exogenous shocks such as changes in the overall interest in new life science technologies. Third, because technology transfer is predicated upon commercialization, a value chain perspective can visualize commercialization obstacles. In this study, a stylized human stem cell therapy value chain is compared to the existing biotechnology value chain and the differences suggest areas where human stem cell therapies are likely to face difficulties. The discussion and conclusion evaluates the progress of the California stem cell initiative and suggests that the evaluation methodology developed can improve the evaluation and guidance of science-based economic development programs.

Susceptibility-Weighted Imaging Identifies Iron-Oxide-Labeled Human Neural Stem Cells: Automated Computational Detection

Neonatal hypoxic-ischemic brain injury (HII) can lead to devastating neurological outcomes such as cerebral palsy, epilepsy, and mental retardation. Human neural stem cell (hNSC) therapy provides new hope for the treatment of neonatal HII. These multipotent cells can aid in HII recovery by activating multiple reparative mechanisms including secretion of neurotrophic factors that enhance brain repair and plasticity. For clinical use of implanted hNSCs, methods are required to identify, quantify, track, and visualize migration and replication in an automated and reproducible fashion. In the current study, we used a model of unilateral HII in 10-day-old rat pups that were implanted with 250,000 Feridex-labeled hNSCs into the contralateral ventricle 3 days after HII. In addition to standard noninvasively acquired serial magnetic resonance imaging (MRI) sequences (11.7 and 4.7 T) that included diffusion-weighted imaging and T2-weighted imaging, we also acquired susceptibility-weighted imaging (SWI) 1-90 days after hNSC implantation. SWI is an advanced MRI method that enhances the visualization of iron-oxide-labeled hNSCs within affected regions of the injured neonatal brain. hNSC contrast was further enhanced by creating minimal intensity projections from the raw SWI magnitude images combined with phase information. Automated computational analysis using hierarchical region splitting (HRS) was applied for semiautomatic detection of hNSCs from SWI images. We found hNSCs in the ipsilateral HII lesion within the striatum and cortex adjacent to the lesion that corresponded to histological staining for iron. Quantitative phase values (radians) obtained from SWI revealed temporally evolving increased phase which reflects a decreased iron oxide content that is possibly related to cell division and the replicative capacity of the implanted hNSCs. SWI images also revealed hNSC migration from the contralateral injection site towards the ipsilateral HII lesion. Our results demonstrate that MRI-based SWI can monitor iron-labeled hNSCs in a clinically relevant manner and that automated computational methods such as HRS can rapidly identify iron-oxide-labeled hNSCs.

Hetrombopag, a Thrombopoietin Receptor Agonist, Protects Cardiomyocyte Survival from Oxidative Stress Damage as an Enhancer of Stem Cells.

Current human umbilical cord blood stem cell therapy faces the great challenges, because the stem cells are scarce and cannot survive for a long time. Here we describe how hetrombopag, an orally-active TPO receptor agonists, enhanced ex vivo expansion of human UCB stem cells, and protected cardiac myocytes from the damage caused by oxidative stress. Ex vivo expansion of stem cells were performed in serum-free medium supplemented with rhSCF and rhFL plus hetrombopag for 7days. The percentage and number of stem cell subsets were determined by flow cytometry. Rat cardiac myocytes, ex vivo expanded stem cells, or cardiac myocytes plus ex vivo expanded stem cells were serum starved for 24hours, and were then subjected to H2O2, hetrombopag or both for 12hours at the indicated concentrations. Cell viability assays, protein microarrays and western blots were then performed in each group. Our studies first revealed that the combination of hetrombopag and rhTPO manifested additive effect on ex vivo expansion of human UCB stem cells. Besides, hetrombopag dose-dependently enhanced the beneficial effects of ex vivo expanded human UCB MNCs in increasing the survival of injured cardiomyocytes during free oxygen radical stress. These data, for the first time, uncovered a novel function of non-peptide small molecular TPO receptor agonists as enhancers of stem cells in protecting cardiac myocyte survival from oxidative stress damage, which might provide a new therapeutic avenue for the treatment of oxidative stress-related cardiovascular disease. Graphical abstract ᅟ.

A Prospective, Randomized, Masked, and Placebo-Controlled Efficacy Study of Intraarticular Allogeneic Adipose Stem Cells for the Treatment of Osteoarthritis in Dogs.

Osteoarthritis (OA) is a degenerative joint disease with a high prevalence in dogs. Mesenchymal stem cells (MSCs) have been used to treat humans, dogs, and horses with OA. This report describes a prospective, randomized, blinded, and placebo-controlled clinical efficacy study of intraarticular allogeneic adipose stem cells for the treatment of dogs with OA. Health assessments and measurements of pain and activity impairment were performed at baseline and at selected time points through day 60. The primary outcome variable was the owner Client-Specific Outcome Measurement (CSOM) and secondary measures included veterinary pain on manipulation, veterinary global score, and owner global score. The dogs were treated with either a saline placebo or a single dose of allogeneic adipose-derived MSCs in either one or two joints. Seventy-four dogs were statistically analyzed for efficacy outcomes. Success in the primary outcome variable, CSOM, was statistically improved in the treated dogs compared to the placebo dogs (79.2 versus 55.4%, p = 0.029). The veterinary pain on manipulation score (92.8 versus 50.2%, p = 0.017) and the veterinary global score (86.9 versus 30.8%, p = 0.009) were both statistically improved in treated dogs compared to placebo. There was no detected significant difference between treated and placebo dogs in the incidence of adverse events or negative health findings. Allogeneic adipose-derived stem cell treatment was shown to be efficacious compared to placebo. This large study of dogs also provides valuable animal clinical safety and efficacy outcome data to our colleagues developing human stem cell therapy.

Label-free biomarkers of human embryonic stem cell differentiation to hepatocytes.

Four different label-free, minimally invasive, live single cell analysis techniques were applied in a quantitative comparison, to characterize embryonic stem cells and the hepatocytes into which they were differentiated. Atomic force microscopy measures the cell's mechanical properties, Raman spectroscopy measures its chemical properties, and dielectrophoresis measures the membrane's capacitance. They were able to assign cell type of individual cells with accuracies of 91% (atomic force microscopy), 95.5% (Raman spectroscopy), and 72% (dielectrophoresis). In addition, stimulated Raman scattering (SRS) microscopy was able to easily identify hepatocytes in images by the presence of lipid droplets. These techniques, used either independently or in combination, offer label-free methods to study individual living cells. Although these minimally invasive biomarkers can be applied to sense phenotypical or environmental changes to cells, these techniques have most potential in human stem cell therapies where the use of traditional biomarkers is best avoided. Destructive assays consume valuable stem cells and do not characterize the cells which go on to be used in therapies; whereas immunolabeling risks altering cell behavior. It was suggested how these four minimally invasive methods could be applied to cell culture, and how they could in future be combined into one microfluidic chip for cell sorting. © 2016 International Society for Advancement of Cytometry.

Enhancing the effectiveness of human cardiac stem cell therapy with the HO-1 inducer-Cobalt Protoporphyrin (CoPP)

Enhancing the effectiveness of human cardiac stem cell therapy with the HO-1 inducer-Cobalt Protoporphyrin (CoPP)