Background
Mesenchymal Stem Cells (MSCs), regardless of the tissue sources, are considered as excellent candidates for cellular therapy as they are immune-privileged cells containing a multitude of ...therapeutic functions that aid in tissue regeneration and repair. For the effective application of these cells in cell therapy, it is important to understand and characterize their biological functions.
Objectives
The present study attempts to characterize the variations in multipotent function such as cell surface antigen levels, proliferation, differentiation and stemness (pluripotency) potential of MSCs isolated from foetal wharton’s jelly (WJ), foetal and maternal side of placenta (PF and PM) and adult tissue sources bone marrow (BM) and adipose tissue (AT) using gene expression by real time PCR (qRT-PCR).
Results
Amongst the different tissue sources, PM, PF and AT-MSCs exhibited significant increase (p < 0.001, p < 0.001 and p < 0.01 respectively) in CD 73 expression and therefore could have a role in immunomodulation. WJ-MSCs exhibited superior proliferation potential based on growth curve, PCNA and Wnt gene expression. BM-MSCs were superior in exhibiting trilineage differentiation. Enhanced stemness potential (Oct 4 and Nanog) was observed for both BM and WJ-MSCs. In addition, BM and WJ-MSCs expressed high levels of CD 90 making them suitable in bone repair and regeneration.
Conclusion
Thus to conclude, out of the five different sources tested, BM an adult source and WJ-MSCs a foetal source were superior in exhibiting most of the biological functions indicating that these sources may be suitable candidates for cell repair and regeneration studies.
Background
Mammosphere formation assay has become a versatile tool to quantify the activity of putative breast cancer stem cells in non-adherent in vitro cultures. However, optimizing the suspension ...culture system is crucial to establish mammosphere cultures from primary breast tumors.
Methods
This study aimed at determining the self-renewal and sphere-forming potential of breast cancer stem-like cells derived from human primary invasive ductal carcinoma and normal breast tissue samples, and MCF-7 breast cancer cell line using an optimal suspension culture system. Mammosphere-forming efficiency of the mammospheres generated from the tissue samples and cell line were compared. We evaluated the expression of CD44
+
/CD24
−
/
low
and CD49f
+
/EpCAM
−
/
low
phenotypes in the stem-like cells by flow cytometry. CK-18, CK-19, α-SMA, and EpCAM marker expression was assessed using immunohistochemical staining.
Results
Breast epithelial cells isolated from the three samples formed two-dimensional spheroids in suspension cultures. Interestingly, mammospheres formed from patient-derived primary breast tumors were enriched in breast cancer stem-like cells with the phenotype CD44
+
/CD24
−
/
low
and exhibited a relatively more number of large spheres when compared to the normal breast stem cells. MCF-7-derived SCs were more aggressive and resulted in the formation of a significantly higher number of spheroids. The expression of CK-18/CK-19 and α-SMA/EpCAM proteins was confirmed in breast cancer tissues.
Conclusions
Thus, the use of primary tumor specimens and breast cancer cell lines as suitable models for elucidating the breast cancer stem cell activity was validated using mammosphere culture system.
Mesenchymal stem cells (MSCs) are highly preferred in clinical therapy for repair and regeneration of diseased tissues for their multipotent properties. Conventionally, MSCs have been cultured in ...media supplemented with animal derived serum, however, it is ideal to expand MSCs in media containing supplements of human origin for clinical therapy. Currently, a number of human derived products are being studied as an alternative to animal sources. Amongst these, platelet lysate (PL) has gained interest in the culture of MSCs without affecting their phenotypic property.
In this study, we used various concentration of PL (2.5, 5, 7.5 & 10%) in the growth medium of MSCs to identify the least concentration of PL that could be an effective alternative to animal products.
MSCs were isolated from Wharton's Jelly by using explant method and expanded in various concentration of PL supplemented medium against the standard FBS containing medium. WJ-MSCs were characterised as per the minimal criteria proposed by International Society for Cell therapy (ISCT), Proliferation study by BrdU assay, gene expression study by qRT-PCR, sterility test for bacteria, Mycoplasma by PCR and endotoxin detection by LAL assay.
Whartons jelly derived MSCs (WJ-MSCs) cultured using standard medium supplemented with various concentration of PL exhibited enhanced proliferation and differentiation potential, unaltered immunophenotypic property and genetic stability when compared with the commercial medium containing 10% FBS.
The least concentration of PL for an ideal expansion of MSCs was found to be 2.5% and was comparable to FBS.
The multipotent and immunosuppressive capacities of mesenchymal stem cells (MSCs) attract several scientists worldwide towards translational research focusing on treatment of diseases including liver ...failure. Though MSC’s have been isolated from different sources, researchers do not concur on the best source for expansion and clinical translation. In this study, we have compared the isolation, proliferation and expansion of MSCs from umbilical cord blood (UCB), Wharton’s Jelly (WJ), bone marrow (BM) and adipose tissue (AT). MSCs were isolated by density gradient separation from UCB, BM and AT and by both enzymatic and explant method for WJ. The MSCs are characterized by their ability to adhere to plastic, expression of positive (CD105, CD73, CD90, CD29, CD44) and negative (CD45, CD14, CD34) markers by flow cytometry and also by their in vitro adipogenic, osteogenic and chondrogenic differentiation. This comprehensive study clearly shows that WJ is better than UCB both in terms of rapidity, yield and ease of procedure. AT and BM are autologous sources for MSC’s but the specimen collection involves cumbersome and painful procedures and an invasive approach. However being autologous, they are safe and probable candidates for therapeutic future applications.
Cell-derived matrices (CDMs) are scaffolds constructed by decellularization of cellular matrices from different tissues and organs. Since CDMs mimic the extracellular matrices (ECMs) of native ...tissues, it plays an essential role in the preparation of bioscaffolds. CDM scaffolds from mesenchymal stem cells (MSCs) have been reported to support cell adhesion and proliferation of its own cells. Therefore, in this study we aimed to test if growth of human Wharton's jelly-derived MSCs may be enhanced when cultured on their own CDMs. To do this, MSCs were induced to generate ECM using ascorbic acid. Thus, obtained matrices were decellularized and characterized quantitatively for changes in their biochemical components (total protein, collagen, glycosaminoglycans) and qualitatively for fibronectin, laminin, and collagen (I & IV) by immunostaining. Our results show the retention of essential ECM components in the decellularized WJ-MSC-derived matrix (WJ-CDM). The influence of WJ-CDM on proliferation and differentiation of WJ-MSCs were evaluated by comparing their growth on collagen and fibronectin-only coated plates. A non-coated tissue culture polystyrene plate (TCPS) served as control. Our cell proliferation results show that no significant changes were observed in the proliferation of MSCs when cultured on WJ-CDM as compared to the bio-coated and non-coated cultures. However, gene expression analysis of the differentiation process showed that osteogenic and adipogenic differentiation potential of the WJ-MSCs was significantly increased upon culturing them on WJ-CDM. In conclusion, the present study reveals that the WJ-MSCs cultured on WJ-CDM may augment osteogenic and adipogenic differentiation.
Objective: Urinary tract infection (UTI) is one of the most common infections observed in diabetic patients. This study is aimed at identifying the organisms with their anti-bacterial resistance ...pattern.
Methods: A total of 400 diabetic patients over a period of nine months presenting with symptom s of urinary tract infection were taken for the study. Their urine were cultured and an antibiogram done.
Results: E. coli, Klebsiella and Enterococci were the commonest organism found. It was found that E. coli, which was the commonest organism E. Coli was sensitive to Norfloxacin and resistant to Ciprofloxacin.
Conclusion: Empirical treatment with ciprofloxacin, Which is considered the drug of choice, will lead to failure of treatment.
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•HSP70, HSP90, NFκB, and BRCA1 play a crucial role in the regulation and sustenance of cancer stem cells.•Anticancer inhibitory effect of withaferin A and its derivatives were ...evaluated against multiple targets.•Molecular docking approach was employed to identify potential natural anti-cancer compounds.•Withaferin A and withaferin A diacetate showed strong inhibitory effects against breast cancer stem-like cells.•Withaferin A and its analogs could be novel anti-cancer agents against breast cancer.
Cancer stem cells (CSCs) in breast cancers are considered to be a major cause of tumor recurrence and metastasis, which lead to chemotherapeutic failures and poor survival rates among breast cancer patients. Hence, this study attempted to evaluate the anti-tumor potential of a bioactive withanolide compound, withaferin A (WFA), against the molecular targets that maintain the stemness of breast cancer stem cells, using in silico molecular modeling approach. The crystal structures of potential breast cancer stem cell targets—HSP70, HSP90, NFκB, and BRCA1 were retrieved from PDB. The 2D structures of WFA, its analogs, along with doxorubicin and vinblastine were retrieved from the PubChem database. WFA and withaferin A diacetate exhibited strong receptor-ligand interactions for the target macromolecules. Their binding energy values ranged between –55.241 and –40.250 kcal/mol. Compared to doxorubicin, WFA markedly inhibited the growth and decreased the viability of MCF-7-derived breast cancer stem cells, with an IC50 value of 1.476 μM for 48 h. Therefore, our results indicate that WFA and its derivatives could be promising anti-tumor agents against breast cancer stem cells. Further in vitro studies are warranted in order to gain a greater understanding of the action of this compound in a physiological setting.
A new pyrrolidine alkaloid named Punigratane was isolated from the rind of Punica granatum. This is the first report of a pyrrolidine-like structure from the rind. The activity of this compound was ...tested in a representative MDR Klebsiella pneumoniae strain which exhibited high efflux pump activity. At a concentration of 6 mg, this compound Punigratane was found to have efflux inhibition activity.
Objective: To compare the antibacterial activity of prosopisjuliflora seed extract against aerobic and anaerobic bacteria.
Methods: A cross sectional study was conducted for 6 mo in the clinical ...microbiology laboratory of SMCH. Agar cup diffusion technique is used to isolate the strains of Clostridium perfringens ATCC 3624, Staphylococcus aureus ATCC 25923 and Escherechiacoli ATCC 25922. The prosopisjulifloraseeds are collected from the saveetha medical college.
Results: By performing the research with proper guidance it is observed that all the three bacterias–Staphylococcus aureus, Escherechia coli, Clostridium perfringens showed sensitivity to prosopisjulifloraseed extract.
Conclusion: Due to its vast antibacterial activity it can be used along with other antibiotics to increase its efficacy. This is used for the treatment of infectious diseases.
Objective: To find out the prevalence and type of microorganisms isolated from mobile phones used by health care workers, students working/studying in a tertiary care center as well as to find the ...rate of contamination of the hands of the individual.
Methods: Swabs moistened with sterile saline was used to swab on phone surfaces and was incubated using standard culture and identification methods. The respective user was instructed to imprint their fingers of both hands on plates of culture media. These were incubated and processed as per standard culture methods.
Results: The most common isolated microorganisms in both groups were Coagulase-negative Staphylococci (CoNS) and MSSA. Among Mobile phones of HCW, the highest contamination rate was noted in physicians 70% followed by Intensive care doctors 60%, and Nurses. Finger impression growth rate was observed high among Nurses 70% followed by Intensive care doctors 60% and physicians 40%.
Conclusion: There is found to be a moderate contamination rate of mobile phones and fingers with pathogenic bacteria as well as normal flora of skin isolated from health care workers. Mobile phones and hands of Health care workers serve as a potential reservoir for hospital-acquired infections as multi-drug resistant pathogenic bacteria. In order to reduce the incidence of nosocomial infections, there should be an implementation of handwashing practices.
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