The goal of this study was to compare the growth and differentiation potential of rhesus monkey mesenchymal stem cells (rhMSCs) from different age groups (fetal, newborn, infant, juvenile), and to use confocal micro-Raman spectroscopy to assess the intrinsic biomolecular profiles of individual rhMSCs. Results indicated that fetal cells had significantly shorter population doubling times during the log growth phase (23.3 +/- 1.3 h) and greater population doubling times (66.5 +/- 6.5) when compared to other age groups (newborn 51.9 +/- 2.3, infant 38.2 +/- 3.1, juvenile 40.7 +/- 4.1). Fetal rhMSCs also differentiated toward osteogenic and adipogenic lineages at a faster rate when compared to cells from older animals. The Raman spectral analysis showed greater DNA and lower protein concentration in fetal compared to juvenile rhMSCs, although the spectra from different age groups shared many similar features. Additionally, principal component analysis (PCA), which is used to discriminate between rhMSCs, supported prior findings that suggested that cultured rhMSCs consist of a heterogeneous cell population. Although the growth potential of rhMSCs from the younger age groups was confirmed, further studies will be necessary to fully explore the potential usefulness of Raman micro-spectroscopy to characterize stem and progenitor cells such as rhMSCs.
"Cells were seeded and grown to approximately 70-80% confluence in 6-well culture plates. To initiate trilineage differentiation, complete DMEM⁄ F12 medium was removed, and tissue-specific differentiation medium was added and maintained for three weeks as described [21-23]. Concretely, osteogenic induction medium (Hyclone) was added to induce osteogenic differentiation and subsequently changed every 3-4 days. "
[Show abstract][Hide abstract] ABSTRACT: Mesenchymal stem cells (MSCs) can be induced to differentiate into neuronal cells under appropriate cellular conditions and transplanted in brain injury and neurodegenerative diseases animal models for neuroregeneration studies. In contrast to the embryonic stem cells (ESCs), MSCs are easily subject to aging and senescence because of their finite ability of self-renewal. MSCs senescence seriously affected theirs application prospects as a promising tool for cell-based regenerative medicine and tissue engineering. In the present study, we established a reversible immortalized mesenchymal stem cells (IMSCs) line by using SSR#69 retrovirus expressing simian virus 40 large T (SV40T) antigen as an alternative to primary MSCs.
The retroviral vector SSR#69 expressing simian virus 40 large T (SV40T) antigen was used to construct IMSCs. IMSCs were identified by flow cytometry to detect cell surface makers. To investigate proliferation and differentiation potential of IMSCs, cell growth curve determination and mesodermal trilineage differentiation tests were performed. Neuronal differentiation characteristics of IMSCs were detected in vitro. Before IMSCs transplantation, we excluded its tumorigenicity in nude mice firstly. The Morris water maze tests and shuttle box tests were performed five weeks after HIBD models received cells transplantation therapy.
In this study, reversible IMSCs were constructed successfully and had the similar morphology and cell surface makers as primary MSCs. IMSCs possessed better ability of proliferation and anti-senescence compared with primary MSCs, while maintained multilineage differentiation capacity. Neural-like cells derived from IMSCs had similar expressions of neural-specific genes, protein expression patterns and resting membrane potential (RMP) compared with their counterparts derived from primary MSCs. There was no bump formation in nude mice subcutaneously injected with IMSCs. IMSCs played same role as primary MSCs to improve learning ability and spatial memory of HIBD rats.
IMSCs not only retain their features of primary MSCs but also possess the ability of high proliferation and anti-senescence. IMSCs can definitely be induced to differentiate into neuronal cells in vitro and take the place of primary MSCs for cell transplantation therapy without tumorigenesis in vivo. The stable cell line is particularly useful and valuable as an alternative to MSCs in neuronal differentiation and neuroregeneration associated studies.
[Show abstract][Hide abstract] ABSTRACT: Many systemic diseases like diabetes, hypertension, or microorganism infections and accident injuries may damage the eyes. Using specific light to enter eyes from the controlled environment, ophthalmologists use light-transmitted information obtained with special instruments like slit-lamp biomicroscopes for clinical diagnosis. In this review, the major structures of the eyes and their important functions will be introduced first. A brief picture of eye diseases is presented, including infectious, immune-related, congenital, degenerative, and traumatic entities. We finally focus on Raman spectroscopy, which is a novel qualitative and quantitative optical technique to probe biological systems with advantages including noninvasiveness, less sample volume needed, and utility even in aqueous environments. The purpose of this paper is to review the applications of this developing non-invasive technology in ophthalmological studies, especially in the diagnostic technology of microbial keratitis, which is a vision-threatening disease necessitating rapid and correct diagnosis. Additionally, some clinical approaches for diagnosing microbial keratitis will also be addressed to fill the gap between clinical methods and this novel technique. Raman-based analysis platform, a promising technique for microbial keratitis diagnosis, can bring a new era, especially for patients with microbial keratitis.
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