Functionalized Scaffold for in Situ Efficient Gene Transfection of Mesenchymal Stem Cells Spheroids toward Chondrogenesis

2018 ◽  
Vol 10 (40) ◽  
pp. 33993-34004 ◽  
Author(s):  
Kunxi Zhang ◽  
Haowei Fang ◽  
Yechi Qin ◽  
Lili Zhang ◽  
Jingbo Yin
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Gene Transfection ◽  
Efficient Gene

Mesenchymal stem cells immortalization's and state-of-art: a systematic review and meta-analysis

2021 ◽  
Vol 31 (Supplement_2) ◽  
Author(s):  
P E F Stricker ◽  
A C Irioda ◽  
B F Mogharbel ◽  
E Abdelwaid ◽  
L R Cavalli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Gene Transfection ◽  
Meta Analysis ◽  
Total N ◽  
Htert Gene ◽  
Cell Immortalization ◽  
Immortalized Cell ◽  
Experimental Strategies ◽  
Established Technique

Abstract Background The immortalization of mesenchymal stem cells (MSCs) allows them to avoid senescence and be cultured through limitless cell passages. Thus, several experimental strategies, such as retrovirus-mediated gene transfer or viral oncogenesis, have been applied for the immortalization of MSCs. The aim was to identifier the most commonly used methodologies and their particularities for the immortalization of human and animal MSCs. Methods The search was conducted in June 2019 and developed in SCOPUS, PUBMED, and SCIENCE DIRECT. Statistical analysis was performed, obtaining the values of total n, mean and standard deviation, confidence interval (CI), and percentage (frequency) for all the predictors. Results The most used immortalization methodology was viral transfection, being the most common immortalized cell type was the bone marrow-derived MSC, and the most used gene for immortalizing both human and animal MSCs was hTERT (39.3%) and SV40T (54.5%). Among the articles analyzed in this review, only 39.3% and 36.4% of human and animal MSCs immortalization protocols, respectively, underwent the tumorigenicity test. Conclusions The virus-mediated gene transfection was observed as the most used and established technique. The insertion of the hTERT gene is still the most used gene for cell immortalization, suggesting that the maintenance of telomerase is efficient for maintaining cell proliferation and bypassing cell senescence. The review concluded that the tumorigenicity tests should become mandatory in order to safely use the immortalized MSCs for translation.

scholarly journals Deciphering the in vivo Dynamic Proteomics of Mesenchymal Stem Cells in Critical Limb Ischemia

2021 ◽  
Vol 9 ◽  
Author(s):  
Yipeng Du ◽  
Xiaoting Li ◽  
Wenying Yan ◽  
Zhaohua Zeng ◽  
Dunzheng Han ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Protein Synthesis ◽  
Critical Limb Ischemia ◽  
Click Reaction ◽  
Limb Ischemia ◽  
Trna Synthetase ◽  
Doppler Imaging

ObjectiveRegenerative therapy using mesenchymal stem cells (MSC) is a promising therapeutic method for critical limb ischemia (CLI). To understand how the cells are involved in the regenerative process of limb ischemia locally, we proposed a metabolic protein labeling method to label cell proteomes in situ and then decipher the proteome dynamics of MSCs in ischemic hind limb.Methods and ResultsIn this study, we overexpressed mutant methionyl-tRNA synthetase (MetRS), which could utilize azidonorleucine (ANL) instead of methionine (Met) during protein synthesis in MSCs. Fluorescent non-canonical amino-acid tagging (FUNCAT) was performed to detect the utilization of ANL in mutant MSCs. Mice with hindlimb ischemia (HLI) or Sham surgery were treated with MetRSmut MSCs or PBS, followed by i.p. administration of ANL at days 0, 2 6, and 13 after surgery. FUNCAT was also performed in hindlimb tissue sections to demonstrate the incorporation of ANL in transplanted cells in situ. At days 1, 3, 7, and 14 after the surgery, laser doppler imaging were performed to detect the blood reperfusion of ischemic limbs. Ischemic tissues were also collected at these four time points for histological analysis including HE staining and vessel staining, and processed for click reaction based protein enrichment followed by mass spectrometry and bioinformatics analysis. The MetRSmut MSCs showed strong green signal in cell culture and in HLI muscles as well, indicating efficient incorporation of ANL in nascent protein synthesis. By 14 days post-treatment, MSCs significantly increased blood reperfusion and vessel density, while reducing inflammation in HLI model compared to PBS. Proteins enriched by click reaction were distinctive in the HLI group vs. the Sham group. 34, 31, 49, and 26 proteins were significantly up-regulated whereas 28, 32, 62, and 27 proteins were significantly down-regulated in HLI vs. Sham at days 1, 3, 7, and 14, respectively. The differentially expressed proteins were more pronounced in the pathways of apoptosis and energy metabolism.ConclusionIn conclusion, mutant MetRS allows efficient and specific identification of dynamic cell proteomics in situ, which reflect the functions and adaptive changes of MSCs that may be leveraged to understand and improve stem cell therapy in critical limb ischemia.

Efficient Gene Delivery to Mesenchymal Stem Cells by an Ethylenediamine-Modified Polysaccharide from Mulberry Leaves

Small ◽  
2011 ◽  
Vol 8 (3) ◽  
pp. 441-451 ◽  
Author(s):  
Wen-Wen Deng ◽  
Xia Cao ◽  
Miao Wang ◽  
Yan Yang ◽  
Wei-Yan Su ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Gene Delivery ◽  
Mulberry Leaves ◽  
Efficient Gene

scholarly journals Preparation of Cationized Polysaccharides as Gene Transfection Carrier for Bone Marrow-Derived Mesenchymal Stem Cells

2010 ◽  
Vol 21 (2) ◽  
pp. 185-204 ◽  
Author(s):  
Jun-ichiro Jo ◽  
Arimichi Okazaki ◽  
Kentaro Nagane ◽  
Masaya Yamamoto ◽  
Yasuhiko Tabata
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Gene Transfection

519 Development and Use of an Intraoperatively Usable Hand-Held Bio-Printer Delivering Mesenchymal Stem Cells In-situ

2019 ◽  
Vol 40 (Supplement_1) ◽  
pp. S235-S236
Author(s):  
G Eylert ◽  
R Cheng ◽  
S He ◽  
J Gariepy ◽  
A Parousis ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells

scholarly journals Gene transfection of human mesenchymal stem cells with a nano-hydroxyapatite-collagen scaffold containing DNA-functionalized calcium phosphate nanoparticles

2016 ◽  
Vol 21 (7) ◽  
pp. 682-695 ◽  
Author(s):  
Taichi Tenkumo ◽  
Juan Ramón Vanegas Sáenz ◽  
Yukyo Takada ◽  
Masatoshi Takahashi ◽  
Olga Rotan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Calcium Phosphate ◽  
Gene Transfection ◽  
Human Mesenchymal Stem Cells ◽  
Collagen Scaffold ◽  
Calcium Phosphate Nanoparticles
Sign in / Sign up

Export Citation Format

Share Document