scholarly journals Monitoring stem cell differentiation using Raman microspectroscopy: chondrogenic differentiation, towards cartilage formation

The Analyst ◽  
2021 ◽  
Vol 146 (1) ◽  
pp. 322-337
Author(s):  
Francesca Ravera ◽  
Esen Efeoglu ◽  
Hugh J. Byrne

Raman microspectroscopy is employed to monitor the differentiation of mesenchymal stem cells to chondrocytes, from subcellular to extracellular matrix evolution.

2021 ◽  
Author(s):  
Anggraini Barlian ◽  
Rizka Musdalifah Amsar ◽  
Salindri Prawitasari ◽  
Christofora Hanny Wijaya ◽  
Ika Dewi Ana ◽  
...  

Abstract BackgroundCells produce extracellular vesicles, such as exosomes and microvesicles, which are used for intracellular communication. Cell-free therapies could be enhanced by using mesenchymal stem cell-derived exosomes. Preconditioning parental cells affects the properties of their exosomes. This study aimed to investigate the role of L-ascorbic acid (LAA) and CoCl2 in the exosomes produced by human Wharton’s jelly mesenchymal stem cells (hWJ MSC) and its potential to induce chondrogenic differentiation of stem cells was also studied.. MethodThe cells were obtained from umbilical cords and characterized based on mesenchymal stem cell criteria. The cells were cultured in a serum-free medium containing LAA and CoCl2. Exosomes produced by the cells were isolated and their morphology observed with Transmission Electron Microscopy. The presence of CD 63 was confirmed using ELISA. The particle size distribution and exosome concentration were analyzed with Nanoparticle Tracking Analysis (NTA). The ability of exosomes to induce stem cell differentiation into chondrocytes was investigated using the Alcian blue assay and immunocytochemistry.ResultsStem cells were successfully isolated from the human umbilical cord. The cells can differentiate into adipocytes, chondrocytes, and osteocytes. Flowcytometry analysis showed the specific surface marker of mesenchymal stem cells. Exosomes isolated from pretreatment cells showed round-shaped morphology and confirmed the presence of CD 63. NTA analysis revealed that pretreatment of cells with LAA increases exosome yields. LAA supplementation in cell medium under hypoxic conditions induced by CoCl2 produces exosomes that can induce the chondrogeic differentiation of stem cells, confirmed by the presence of glycosaminoglycan and collagen type 2.ConclusionExosomes produced by preconditioning hWJ-MSC with LAA in hypoxic conditions have the potential to enhance human Wharton Jelly stem cell differentiation into chondrocytes.


2015 ◽  
Vol 35 (10) ◽  
pp. 1700-1711 ◽  
Author(s):  
Fenfang Chen ◽  
Xia Lin ◽  
Pinglong Xu ◽  
Zhengmao Zhang ◽  
Yanzhen Chen ◽  
...  

Bone morphogenetic proteins (BMPs) play vital roles in regulating stem cell maintenance and differentiation. BMPs can induce osteogenesis and inhibit myogenesis of mesenchymal stem cells. Canonical BMP signaling is stringently controlled through reversible phosphorylation and nucleocytoplasmic shuttling of Smad1, Smad5, and Smad8 (Smad1/5/8). However, how the nuclear export of Smad1/5/8 is regulated remains unclear. Here we report that the Ran-binding protein RanBP3L acts as a nuclear export factor for Smad1/5/8. RanBP3L directly recognizes dephosphorylated Smad1/5/8 and mediates their nuclear export in a Ran-dependent manner. Increased expression of RanBP3L blocks BMP-induced osteogenesis of mouse bone marrow-derived mesenchymal stem cells and promotes myogenic induction of C2C12 mouse myoblasts, whereas depletion of RanBP3L expression enhances BMP-dependent stem cell differentiation activity and transcriptional responses. In conclusion, our results demonstrate that RanBP3L, as a nuclear exporter for BMP-specific Smads, plays a critical role in terminating BMP signaling and regulating mesenchymal stem cell differentiation.


2015 ◽  
Vol 3 (16) ◽  
pp. 3150-3168 ◽  
Author(s):  
Sunil Kumar Boda ◽  
Greeshma Thrivikraman ◽  
Bikramjit Basu

Substrate magnetization as a tool for modulating the osteogenesis of human mesenchymal stem cells for bone tissue engineering applications.


Physiology ◽  
2018 ◽  
Vol 33 (1) ◽  
pp. 16-25 ◽  
Author(s):  
Lucas R. Smith ◽  
Sangkyun Cho ◽  
Dennis E. Discher

Stem cells mechanosense the stiffness of their microenvironment, which impacts differentiation. Although tissue hydration anti-correlates with stiffness, extracellular matrix (ECM) stiffness is clearly transduced into gene expression via adhesion and cytoskeleton proteins that tune fates. Cytoskeletal reorganization of ECM can create heterogeneity and influence fates, with fibrosis being one extreme.


2016 ◽  
Vol 8 (41) ◽  
pp. 7437-7444 ◽  
Author(s):  
Hongjun Song ◽  
Jenna M. Rosano ◽  
Yi Wang ◽  
Charles J. Garson ◽  
Balabhaskar Prabhakarpandian ◽  
...  

A dual-micropore-based microfluidic electrical impedance flow cytometer for non-invasive identification of the differentiation state of mesenchymal stem cells.


Small ◽  
2016 ◽  
Vol 12 (13) ◽  
pp. 1770-1778 ◽  
Author(s):  
Jichuan Qiu ◽  
Jianhua Li ◽  
Shu Wang ◽  
Baojin Ma ◽  
Shan Zhang ◽  
...  

Author(s):  
Yuxuan Zhong ◽  
Xiang Li ◽  
Fanglin Wang ◽  
Shoushuai Wang ◽  
Xiaohong Wang ◽  
...  

The mesenchymal stem cells have multidirectional differentiation potential and can differentiate into adipocytes, osteoblasts, cartilage tissue, muscle cells and so on. The adipogenic differentiation of mesenchymal stem cells is of great significance for the construction of tissue-engineered fat and the treatment of soft tissue defects. Exosomes are nanoscale vesicles secreted by cells and widely exist in body fluids. They are mainly involved in cell communication processes and transferring cargo contents to recipient cells. In addition, exosomes can also promote tissue and organ regeneration. Recent studies have shown that various exosomes can influence the adipogenic differentiation of stem cells. In this review, the effects of exosomes on stem cell differentiation, especially on adipogenic differentiation, will be discussed, and the mechanisms and conclusions will be drawn. The main purpose of studying the role of these exosomes is to understand more comprehensively the influencing factors existing in the process of stem cell differentiation into adipocytes and provide a new idea in adipose tissue engineering research.


2020 ◽  
Vol 15 (5) ◽  
pp. 462-472 ◽  
Author(s):  
Abdollah Amini ◽  
Sufan Chien ◽  
Mohammad Bayat

Objective: This is a systematic review of the effects of low-intensity pulsed ultrasound (LIPUS) on stem cell differentiation. Background Data: Recent studies have investigated several types of stem cells from different sources in the body. These stem cells should strictly be certified and promoted for cell therapies before being used in medical applications. LIPUS has been used extensively in treatment centers and in research to promote stem cell differentiation, function, and proliferation. Materials and Methods: The databases of PubMed, Google Scholar, and Scopus were searched for abstracts and full-text scientific papers published from 1989-2019 that reported the application of LIPUS on stem cell differentiation. Related English language articles were found using the following defined keywords: low-intensity pulsed ultrasound, stem cell, differentiation. Criteria for inclusion in the review were: LIPUS with frequencies of 1–3 MHz and pulsed ultrasound intensity of <500 mW/cm2. Duration, exposure time, and cell sources were taken into consideration. Results: Fifty-two articles were selected based on the inclusion criteria. Most articles demonstrated that the application of LIPUS had positive effects on stem cell differentiation. However, some authors recommended that LIPUS combined with other physical therapy aides was more effective in stem cell differentiation. Conclusions: LIPUS significantly increases the level of stem cell differentiation in cells derived mainly from bone marrow mesenchymal stem cells. There is a need for further studies to analyze the effect of LIPUS on cells derived from other sources, particularly adipose tissue-derived mesenchymal stem cells, for treating hard diseases, such as osteoporosis and diabetic foot ulcer. Due to a lack of reporting on standard LIPUS parameters in the field, more experiments comparing the protocols for standardization of LIPUS parameters are needed to establish the best protocol, which would allow for the best results.


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