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2022 ◽  
Author(s):  
Joo Youn Lee ◽  
Min Hee Kang ◽  
Ji Eun Jang ◽  
Jeong Eon Lee ◽  
Yuyeong Yang ◽  
...  

Abstract Stem cells are attractive candidates for the regeneration of tissue and organ. Mesenchymal stem cells (MSCs) have been extensively investigated for their potential applications in regenerative medicine and cell therapy. For developing effective stem cell therapy, the mass production of consistent quality cells is required. The cell culture medium is the most critical aspect of the mass production of qualified stem cells. Classically, fetal bovine serum (FBS) has been used as a culture supplement for MSCs. Due to the undefined and heterologous composition of animal origin components in FBS, efforts to replace animal-derived components with non-animal-derived substances led to safe serum free media (SFM). Adipose derived mesenchymal stem cells (ADSCs) cultivated in SFM provided a more stable population doubling time (PDT) to later passage and more cells in a shorter time compared to FBS containing media. ADSCs cultivated in SFM had lower cellular senescence, lower immunogenicity, and higher genetic stability than ADSCs cultivated in FBS containing media. Differential expression analysis of mRNAs and proteins showed that the expression of genes related with apoptosis, immune response, and inflammatory response were significantly up-regulated in ADSCs cultivated in FBS containing media. ADSCs cultivated in SFM showed similar therapeutic efficacy in an acute pancreatitis mouse model to ADSCs cultivated in FBS containing media. Consideration of clinical trials, not only pre-clinical trial, suggests that cultivation of MSCs using SFM might offer more safe cell therapeutics as well as repeated administration due to low immunogenicity.


2022 ◽  
Author(s):  
Aijia Cai ◽  
Paul Schneider ◽  
Zeng-Ming Zheng ◽  
Justus P. Beier ◽  
Marcus Himmler ◽  
...  

Abstract Primary myoblasts (Mb) and adipose derived mesenchymal stromal cells (ADSC) can be co-cultured and myogenically differentiated in the process of skeletal muscle tissue engineering. Electrospun composite nanofiber scaffolds represent suitable matrices for tissue engineering of skeletal muscle, combining biocompatibility and stability. Although growth differentiation factor 11 (GDF11) has been proposed as a rejuvenating circulating factor, restoring skeletal muscle function in aging mice, some studies have also described a harming effect of GDF11.Therefore the aim of the study was to analyze the effect of GDF11 on co-cultures of Mb and ADSC on poly-ε-caprolacton (PCL)-collagen I-polyethylene oxide (PEO)-nanofibers.Human Mb were co-cultured with ADSC two-dimensionally (2D) as monolayers or three-dimensionally (3D) on aligned PCL-collagen I-PEO-nanofibers. Differentiation media were either serum-free with or without GDF11, or serum containing as in a conventional differentiation medium. Cell viability was higher after conventional myogenic differentiation compared to serum-free and serum-free + GDF11 differentiation as was creatine kinase activity. Immunofluorescence staining showed myosin heavy chain expression in all groups after 28 days of differentiation. Gene expression of myosin heavy chain (MYH2) increased after serum-free + GDF11 stimulation compared to serum-free stimulation alone. The results of this study show that PCL-collagen I-PEO-nanofibers represent a suitable matrix for 3D myogenic differentiation of Mb and ADSC. In this context, GDF11 seems to promote myogenic differentiation of Mb and ADSC co-cultures compared to serum-free differentiation without any evidence of a harming effect.


2021 ◽  
Vol 5 (2) ◽  
pp. 161-167
Author(s):  
O. A. Zhigaltsova-Kuchinskaya ◽  
◽  
N. N. Silivontchik ◽  
S. A. Likhachev ◽  
I. V. Pleshko ◽  
...  

Bacground. The optimization of Wilson’s disease (WD) diagnosis is one of the most disputable problem. Objective. The retrospective study of initial assessment findings under clinical suspicion for WD in 102 patients with the confirmed diagnosis. Material and methods. The results of laboratory tests and Kaiser-Fleischer rings (KF rings) identification under clinical suspicion for WD in 102 patients with the confirmed diagnosis. Results. At stage I, 17 patients (16.7%; 95% CI 10.7–25.1) were defined as having clinically definitive WD based on the combination of low serum ceruloplasmin and KF rings, 4 patients (3.9%; 95% CI 1.5–9.7) – based on the drop of ceruloplasmin level. After stage II, involving 24-hour urinary copper excretion evaluation, the rate of definitive diagnosis of WD reached 24,5% (95% CI 17.2 33.7). After stage III (genotyping for carriage of ATP7B gene mutations) – 56.9% (95% CI 47.2–66.0). Serum free copper increase was found in 54.9% (95% CI 41.4 67.7) of cases. Conclusions. Under clinical suspicion for WD, initial structured ophthalmological, laboratory and molecular-genetic assessment ensured the diagnosis of WD only in 56.9% (95% CI 56.9; 47.2–66.1). Frequent detection of serum free copper increase (54.9%, 95% CI 41.4 67.7) allows to use this test due to its greater availability as compared with 24-hour urinary copper excretion evaluation in WD diagnostics.


Antioxidants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2022
Author(s):  
Larissa E. van Eijk ◽  
Adriana Tami ◽  
Jan-Luuk Hillebrands ◽  
Wilfred F. A. den Dunnen ◽  
Martin H. de Borst ◽  
...  

Oxidative stress has been implicated to play a critical role in the pathophysiology of coronavirus disease 2019 (COVID-19) and may therefore be considered as a relevant therapeutic target. Serum free thiols (R-SH, sulfhydryl groups) comprise a robust marker of systemic oxidative stress, since they are readily oxidized by reactive oxygen species (ROS). In this study, serum free thiol concentrations were measured in hospitalized and non-hospitalized patients with COVID-19 and healthy controls and their associations with relevant clinical parameters were examined. Serum free thiol concentrations were measured colorimetrically (Ellman’s method) in 29 non-hospitalized COVID-19 subjects and 30 age-, sex-, and body-mass index (BMI)-matched healthy controls and analyzed for associations with clinical and biochemical disease parameters. Additional free thiol measurements were performed on seven serum samples from COVID-19 subjects who required hospitalization to examine their correlation with disease severity. Non-hospitalized subjects with COVID-19 had significantly lower concentrations of serum free thiols compared to healthy controls (p = 0.014), indicating oxidative stress. Serum free thiols were positively associated with albumin (St. β = 0.710, p < 0.001) and inversely associated with CRP (St. β = −0.434, p = 0.027), and showed significant discriminative ability to differentiate subjects with COVID-19 from healthy controls (AUC = 0.69, p = 0.011), which was slightly higher than the discriminative performance of CRP concentrations regarding COVID-19 diagnosis (AUC = 0.66, p = 0.042). This study concludes that systemic oxidative stress is increased in patients with COVID-19 compared with healthy controls. This opens an avenue of treatment options since free thiols are amenable to therapeutic modulation.


2021 ◽  
Author(s):  
Yiqi Yang ◽  
Bihui Zhang ◽  
Junye Xie ◽  
Yuling Cai ◽  
Jia Liu ◽  
...  

Abstract Background: Umbilical cord blood (UCB) has been clinically used for human hematopoietic stem cells (HSCs) transplantation. However, limited numbers of the functional UCB-HSCs from single cord blood restricts its application in adults, while most of the strategies for stem cells expansion in vitro are either inefficient or costly. To overcome these obstacles, we evaluated the potential role of our newly identified CH02 peptide in ex vivo culture expansion of CD34+ UCB-HSCs. Methods: Enriched human CD34+ progenitor/stem cells populations were cultured in serum-free medium supplemented with different cytokines combinations for 8 days. These cytokines combinations included various concentration of CH02 peptide or the FLT3 ligand, with a cocktail of several growth factors such as IL-6, SCF and TPO. In addition, the global gene expression profile of the CD34+ cells cultured under different conditions were monitored through RNA-seq experiments. Furthermore, the expanded CD34+ cells were topically transplanted into the dorsal wounds of diabetic mice, and the wound closure was observed to evaluate the pro-repair ability of CH02-cultured CD34+ cells.Results: We herein report that the combination of CH02 peptide and other cytokines under the serum-free medium can effectively expand the CD34+ HSCs into 12-fold within 7 days while maintaining their stem cell properties. Moreover, CH02 peptide increased the anti-inflammatory and growth-promoting capacity of CD34+ cells, and thus accelerating wound healing of diabetic mice via promoting the anti-inflammatory and inhibiting the inflammatory factors.Conclusions: Together, our CH02 peptide demonstrated promising potentials to improve HSCs expansion for clinical application.


Author(s):  
Chandra Mohan ◽  
Kunal Gururani ◽  
Anurag Rawat ◽  
Mansi Kala

Background: Data on relationship between serum testosterone and endothelial dysfunction measured by brachial artery flow-mediated dilatation (BAFMD) in Indian subset are scarce. The present study was envisaged to assess the correlation between serum testosterone and endothelial dysfunction measured by BAFMD.Methods: From October 2013 till September 2014, 92 Indian male patients aged 40-60 years who underwent investigation of flow-mediated dilatation of the brachial artery using ultra sonography were included. The association between serum testosterone and BAFMD percent-measured endothelial dysfunction was examined.Results: Multivariate regression analysis in 92 Indian male patients (mean age 53.12±6.3 years) revealed that low levels of total serum, serum free and serum bioavailable testosterone were significantly associated with BAFMD% and were independent of age, hypertension, diabetes, body mass index (BMI), current smoking and hyperlipidaemia (p<0.001). The total serum, serum free and serum bioavailable testosterone were positively correlated with BAFMD% with Pearson correlation coefficients of r=0.572, r=0.525 and r=0.547, respectively (p<0.001).Conclusions: Low levels of total serum, serum free and serum bioavailable testosterone were significantly associated with BAFMD%-measured endothelial dysfunction, irrespective of cardiovascular risk factors.


2021 ◽  
Vol 22 (24) ◽  
pp. 13449
Author(s):  
Wei-Wei Gao ◽  
Jie Zheng ◽  
Wonjin Yun ◽  
Phil-Jun Kang ◽  
Gyuman Park ◽  
...  

Background: Regenerative medicine strategies employing nephron progenitor cells (NPCs) are a viable approach that is worthy of substantial consideration as a promising cell source for kidney diseases. However, the generation of induced nephron progenitor-like cells (iNPCs) from human somatic cells remains a major challenge. Here, we describe a novel method for generating NPCs from human urine-derived cells (UCs) that can undergo long-term expansion in a serum-free condition. Results: Here, we generated iNPCs from human urine-derived cells by forced expression of the transcription factors OCT4, SOX2, KLF4, c-MYC, and SLUG, followed by exposure to a cocktail of defined small molecules. These iNPCs resembled human embryonic stem cell-derived NPCs in terms of their morphology, biological characteristics, differentiation potential, and global gene expression and underwent a long-term expansion in serum-free conditions. Conclusion: This study demonstrates that human iNPCs can be readily generated and expanded, which will facilitate their broad applicability in a rapid, efficient, and patient-specific manner, particularly holding the potential as a transplantable cell source for patients with kidney disease.


2021 ◽  
Author(s):  
Kristina Thamm ◽  
Kristin Moebus ◽  
Russel Towers ◽  
Stefan Baertschi ◽  
Richard Wetzel ◽  
...  

Mesenchymal stromal cells (MSCs) are one of the most frequently used cell types in regenerative medicine and cell therapy. Generating sufficient cell numbers for MSC-based therapies is constrained by: 1) their low abundance in tissues of origin, which imposes the need for significant ex vivo cell amplification, 2) donor-specific characteristics including MSC frequency/quality that decline with disease state and increasing age, 3) cellular senescence, which is promoted by extensive cell expansion and results in decreased therapeutic functionality. The final yield of a manufacturing process is therefore primarily determined by the applied isolation procedure and its efficiency in isolating therapeutically active cells from donor tissue. To date, MSCs are predominantly isolated using media supplemented with either serum or its derivatives, which pose safety and consistency issues. To overcome those limitations while enabling robust MSC production with constant high yield and quality, we developed a chemically defined biomimetic surface coating, called isoMATRIX, that facilitates the isolation of significantly higher numbers of MSCs in xeno-/serum-free and chemically defined conditions. The isolated cells display a smaller cell size and higher proliferation rate than those derived from a serum-containing isolation procedure and a strong immunomodulatory capacity. In sum, the isoMATRIX promotes enhanced xeno-, serum-free, or chemically defined isolation of human MSCs and supports consistent and reliable cell performance for improved stem cell-based therapies.


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