Single Cell Analysis Reveals That IL-4 Receptor/Stat6 Signaling Is Not Required for the In Vivo or In Vitro Development of CD4+Lymphocytes with a Th2 Cytokine Profile

The repair of injured tendons remains a formidable clinical challenge because of our limited understanding of tendon stem cells and the regulation of tenogenesis. With single-cell analysis to characterize the gene expression profiles of individual cells isolated from tendon tissue, a subpopulation of nestin+ tendon stem/progenitor cells (TSPCs) was identified within the tendon cell population. Using Gene Expression Omnibus datasets and immunofluorescence assays, we found that nestin expression was activated at specific stages of tendon development. Moreover, isolated nestin+ TSPCs exhibited superior tenogenic capacity compared to nestin− TSPCs. Knockdown of nestin expression in TSPCs suppressed their clonogenic capacity and reduced their tenogenic potential significantly both in vitro and in vivo. Hence, these findings provide new insights into the identification of subpopulations of TSPCs and illustrate the crucial roles of nestin in TSPC fate decisions and phenotype maintenance, which may assist in future therapeutic strategies to treat tendon disease.

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The Trends of Single-Cell Analysis: A Global Study

BioMed Research International ◽

10.1155/2020/7425397 ◽

2020 ◽

Vol 2020 ◽

pp. 1-10

Author(s):

Hua Tian ◽

Haifeng Liu ◽

Yuanyuan Zhu ◽

Dan Xing ◽

Bin Wang

Keyword(s):

Single Cell ◽

Single Cell Analysis ◽

Bibliographic Coupling ◽

In Vivo Studies ◽

Cell Analysis ◽

The Usa ◽

Visualization Of Data ◽

Number Of Publications

Objective. The field of single-cell analysis has rapidly grown worldwide, and a bibliometric analysis and visualization of data and publications pertaining to such single-cell research has the potential to offer insights into the development of this field over the past two decades while also highlighting future avenues of research. Methods. Single-cell analysis-related studies published from 2000-2019 were identified through searches of the Web of Science, Scopus, and PubMed databases, and corresponding bibliometric data were systematically compiled. Extracted data from each study included author names, country of origin, and affiliations. GraphPad Prism was used to analyze these data, while VOSviewer was used to perform global analyses of bibliographic coupling, coauthorship, cocitation, and co-occurrence. Results. In total, 4,071 relevant studies were included in this analysis. The number of publications increased substantially with time, suggesting that single-cell analyses are becoming increasingly more prevalent in recent years. Studies from the USA had the greatest impact in this field, with higher H -index values and numbers of citations relative to other countries, whereas Israel exhibited the highest average number of citations per publication. Bibliographic coupling, coauthorship, cocitation, and co-occurrence analyses revealed that Analytical Chemistry was associated with the highest number of publications in this field, and the University of Stanford contributed the most to this field. The most cited study included in this analysis was published by Macosko et al. in 2015 in Cell. Co-occurrence analyses revealed that the most common single-cell research topics included “mechanistic studies,” “in vitro studies,” “in vivo studies,” and “fabrication studies.” Conclusions. Single-cell analyses are a rapidly growing area of scientific interest, and higher volumes of publications in this field are expected in the coming years, particularly for studies conducting fabrication and in vivo single-cell analyses.

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Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

10.1101/2020.03.04.975177 ◽

2020 ◽

Author(s):

Brian S. Iskra ◽

Logan Davis ◽

Henry E. Miller ◽

Yu-Chiao Chiu ◽

Alexander R. Bishop ◽

...

Keyword(s):

Cell Culture ◽

Single Cell ◽

Single Cell Analysis ◽

Cell Types ◽

Cell Analysis ◽

Mass Cytometry ◽

Single Cell Rna Sequencing ◽

High Depth

AbstractCardiac non-myocytes comprise a diverse and crucial cell population in the heart that plays dynamic roles in cardiac wound healing and growth. Non-myocytes broadly fall into four cell types: endothelium, fibroblasts, leukocytes, and pericytes. Here we characterize the diversity of the non-myocytes in vivo and in vitro using mass cytometry. By leveraging single-cell RNA sequencing we inform the design of a mass cytometry panel. To aid in annotation of the mass cytometry datasets, we utilize data integration with a neural network. We introduce approximately 460,000∼ single cell proteomes of non-myocytes as well as 5,000∼ CD31 negative single cell transcriptomes. Using our data, as well as previously reported datasets, we characterize cardiac non-myocytes with high depth in six mice, characterizing novel surface markers (CD9, CD200, Notch3, and FolR2). Further, we find that extended cell culture promotes the proliferation of CD45+CD11b+FolR2+IAIE- myeloid cells in addition to fibroblasts.

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Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

International Journal of Molecular Sciences ◽

10.3390/ijms22168367 ◽

2021 ◽

Vol 22 (16) ◽

pp. 8367

Author(s):

Hien Lau ◽

Shiri Li ◽

Nicole Corrales ◽

Samuel Rodriguez ◽

Mohammadreza Mohammadi ◽

...

Keyword(s):

Tissue Culture ◽

Oxygen Consumption ◽

Insulin Secretion ◽

Oxygen Consumption Rate ◽

Consumption Rate ◽

In Vitro Development ◽

Porcine Islets ◽

Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

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Fabrication of high-aspect-ratio microstructures in polymer microfluid chips for in vitro single-cell analysis

Technical Physics ◽

10.1134/s106378421610008x ◽

2016 ◽

Vol 61 (10) ◽

pp. 1566-1571 ◽

Cited By ~ 10

Author(s):

A. S. Bukatin ◽

I. S. Mukhin ◽

E. I. Malyshev ◽

I. V. Kukhtevich ◽

A. A. Evstrapov ◽

...

Keyword(s):

Aspect Ratio ◽

Single Cell ◽

High Aspect Ratio ◽

Single Cell Analysis ◽

Cell Analysis

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P-165 Cytometry-Based Single Cell Analysis of Intact Epithelial Signaling Reveals MAPK Activation Divergent from TNF-α-Induced Apoptosis in Vivo

Inflammatory Bowel Diseases ◽

10.1097/01.mib.0000480283.46656.d8 ◽

2016 ◽

Vol 22 ◽

pp. S59-S60

Author(s):

Alan Simmons ◽

Amrita Banerjee ◽

Eliot McKinley ◽

Cherieʼ Scurrah ◽

Jeffrey Franklin ◽

...

Keyword(s):

Single Cell ◽

Single Cell Analysis ◽

Cell Analysis ◽

Mapk Activation ◽

Tnf Α ◽

Induced Apoptosis

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Single cell analysis in Vivo

Optics in the Life Sciences ◽

10.1364/boda.2015.bw1a.1 ◽

2015 ◽

Author(s):

Charles P. Lin

Keyword(s):

Single Cell ◽

Single Cell Analysis ◽

Cell Analysis

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Single-cell analysis reveals IGF-1 potentiation of inhibition of the TGF-β/Smad pathway of fibrosis in human keratocytes in vitro

Scientific Reports ◽

10.1038/srep34373 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 11

Author(s):

Tomislav Sarenac ◽

Martin Trapecar ◽

Lidija Gradisnik ◽

Marjan Slak Rupnik ◽

Dusica Pahor

Keyword(s):

Single Cell ◽

Single Cell Analysis ◽

Cell Analysis ◽

Smad Pathway ◽

Human Keratocytes

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64COMPARISON OF DEVELOPMENTAL POTENTIAL OF IN VIVO AND IN VITRO RECIPIENT OOCYTES AFTER NUCLEAR TRANSFER IN GOAT

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab64 ◽

2004 ◽

Vol 16 (2) ◽

pp. 154

Author(s):

H.S. Park ◽

M.Y. Lee ◽

S.P. Hong ◽

J.I. Jin ◽

J.K. Park ◽

...

Keyword(s):

Nuclear Transfer ◽

Electric Pulse ◽

Serum Starvation ◽

Cleavage Rate ◽

Developmental Potential ◽

In Vitro Development ◽

Significant Difference ◽

Vitro Development

Recent techniques in somatic cell nuclear transfer (SCNT) have been widely used for animal research. In addition, SCNT techniques may allow for the rescue of endangered species. Despite efforts for wildlife preservation, however, some threatened or endangered wild animal species will likely become extinct. As a preliminary experiment of a series in wildlife research, we tried to identify an improved method for the production of more transferable NT embryos in goats. Mature donor animals of Korean native goats (20–25kg) were synchronized with a CIDR (type G; InterAg, New Zealand) vaginal implant for 10 days followed by a total of 8 twice daily injections of 70mg of FSH (Folltropine, London, Ontario, Canada) and 400IU of hCG (Chorulon, Intervet, Moxmeer, The Netherlands). Oocytes were then collected surgically by retograde oviduct flush or direct aspiration from ovarian follicles in vivo at 29–34h after hCG. Oocytes collected from follicles were matured in TCM-199 containing 10% FBS and hormones. Prepared ear skin cells from the goat were cultured in TCM-199 containing 10% FBS at 39°C, 5% CO2 in air, and confluent monolayers were obtained. Oocytes were enucleated and donor cells from serum starvation (0.5%) culture were fused through a single electric pulse (DC 2.36kvcm−1, 17μs), and then activated by a single electric pulse (AC 5vmm−1, 5s+DC 1.56kvcm−1, 30μs) or chemical treatment (5μgmL−1 ionomycin 5min−1, 1.9mM 6-DMAP/4h). Reconstructed oocytes were cultured in M16 medium with 10% goat serum (GS) for 6–7 days. Data were analyzed by chi-square test. In in vitro development, significantly (P<0.05) more oocytes were cleaved (24/30, 80.0%) and developed (7/24, 29.2%) to morula or blastocyst stage, respectively, in NT oocytes activated by Iono + DMAP compared to electric stimulated oocytes (2/21, 40.0%; 0/2, 0%). There was a significant difference in in vitro development of NT embryos by the method of oocyte collection. Cleavage rate was higher (P<0.05) in NT embryos from in vivo oocytes (23/28, 82.1%) than in in vitro matured oocytes (19/35, 54.3%), and further development to morula or blastocyst was also significantly (P<0.05%) higher in NT embryos from in vivo oocytes (7/23, 30.4%) than in NT embryos from in vitro matured oocytes (0/19, 0%). When we compared NT embryos to parthenotes, developmental rate was not significantly different between NT embryos and parthenotes. These results strongly suggest that the in vivo oocytes will have superior developmental potential to oocytes matured in vitro. Table 1 Effect of different oocyte source on in vitro development following caprine SCNT

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