scholarly journals Developing a New Two-Step Protocol to Generate Functional Hepatocytes from Wharton’s Jelly-Derived Mesenchymal Stem Cells under Hypoxic Condition

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Patcharee Prasajak ◽  
Wilairat Leeanansaksiri
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Lines ◽  
High Efficiency ◽  
Hypoxic Condition ◽  
Wharton’S Jelly ◽  
New Method ◽  
Cell Physiology ◽  
Wharton's Jelly ◽  
Cell Based Therapy

The shortage of donor livers and hepatocytes is a major limitation of liver transplantation. Thus, generation of hepatocyte-like cells may provide alternative choice for therapeutic applications. In this study, we developed a new method to establish hepatocytes from Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) cell lines named WJMSCs-SUT1 and WJMSCs-SUT2 under hypoxic condition. This new method could rapidly drive both WJ-MSCs cell lines into hepatic lineage within 18 days. The achievement of hepatogenic differentiation was confirmed by the characterization of both phenotypes and functions. More than 80% MSCs-derived hepatocyte-like cells (MSCDHCs) achieved functional hepatocytes including hepatic marker expressions both at gene and protein levels, glycogen storage, low-density lipoprotein uptake, urea production, and albumin secretion. This study highlights the establishment of new hepatogenic induction protocol under hypoxic condition in order to mimic hypoxic microenvironment in typical cell physiology. In conclusion, we present a simple, high-efficiency, and time saving protocol for the generation of functional hepatocyte-like cells from WJ-MSCs in hypoxic condition. The achievement of this method may overcome the limitation of donor hepatocytes and provides a new avenue for therapeutic value in cell-based therapy for life-threatening liver diseases, regenerative medicine, toxicity testing for pharmacological drug screening, and other medical related applications.

scholarly journals Embryonic stem cells conditioned medium enhances Wharton’s jelly-derived mesenchymal stem cells expansion under hypoxic condition

2014 ◽  
Vol 67 (3) ◽  
pp. 493-505 ◽  
Author(s):  
Patcharee Prasajak ◽  
Piyaporn Rattananinsruang ◽  
Kamonnaree Chotinantakul ◽  
Chavaboon Dechsukhum ◽  
Wilairat Leeanansaksiri
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Embryonic Stem Cells ◽  
Conditioned Medium ◽  
Embryonic Stem ◽  
Hypoxic Condition ◽  
Wharton’S Jelly ◽  
Wharton's Jelly

scholarly journals Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

Reproduction ◽  
2012 ◽  
Vol 143 (4) ◽  
pp. 455-468 ◽  
Author(s):  
Eleonora Iacono ◽  
Lara Brunori ◽  
Alessandro Pirrone ◽  
Pasquale Paolo Pagliaro ◽  
Francesca Ricci ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Amniotic Fluid ◽  
Cord Blood ◽  
Cell Lines ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Wharton’S Jelly ◽  
Blue Staining ◽  
Wharton's Jelly

Mesenchymal stem cells (MSCs) have been derived from multiple sources of the horse including umbilical cord blood (UCB) and amnion. This work aimed to identify and characterize stem cells from equine amniotic fluid (AF), CB and Wharton's Jelly (WJ). Samples were obtained from 13 mares at labour. AF and CB cells were isolated by centrifugation, while WJ was prepared by incubating with an enzymatic solution for 2 h. All cell lines were cultured in DMEM/TCM199 plus fetal bovine serum. Fibroblast-like cells were observed in 7/10 (70%) AF, 6/8 (75%) CB and 8/12 (66.7%) WJ samples. Statistically significant differences were found between cell-doubling times (DTs): cells isolated from WJ expanded more rapidly (2.0±0.6 days) than those isolated from CB (2.6±1.3 days) and AF (2.3±1.0 days) (P<0.05). Positive von Kossa and Alizarin Red S staining confirmed osteogenesis. Alcian Blue staining of matrix glycosaminoglycans illustrated chondrogenesis and positive Oil Red O lipid droplets staining suggested adipogenesis. All cell lines isolated were positive for CD90, CD44, CD105; and negative for CD34, CD14 and CD45. These findings suggest that equine MSCs from AF, UCB and WJ appeared to be a readily obtainable and highly proliferative cell lines from a uninvasive source that may represent a good model system for stem cell biology and cellular therapy applications in horses. However, to assess their use as an allogenic cell source, further studies are needed for evaluating the expression of markers related to cell immunogenicity.

scholarly journals Hypoxic and Normoxic-Human Wharton’s Jelly Mesenchymal Stem Cell-Free Lysate for Anticancer Therapies

2021 ◽  
Vol 18 (9) ◽  
Author(s):  
Wahyu WIDOWATI ◽  
Diana Krisanti JASAPUTRA ◽  
Hanna Sari Widya KUSUMA ◽  
Rizal RIZAL ◽  
Dian Ratih LAKSMITAWATI ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Mouse Fibroblast ◽  
Wharton’S Jelly ◽  
Fibroblast Cells ◽  
Normal Cells ◽  
Wharton's Jelly

Cancer can be defined as a disease in which a group of uncontrolled cells growth by ignoring the rules of normal cell division. Mesenchymal Stem Cells (MSCs) are one of the promising cell-based therapies for anticancer therapies. Preceding research had disclosed that conditioned medium from hypoxic-treated human Wharton’s jelly mesenchymal stem cells (hWJMSCs-hypoCM) and normoxic-treated hWJMSCs (hWJMSCs-norCM) can inhibit the proliferation of various cancer cell lines and have no cytotoxic effect on normal cells. The present study, hence, measured the effect of a human Wharton’s jelly stem cells cell-free lysate (hWJMSCs-CL), cultured with hypoxic condition (hWJMSCs-hypoCL), and normoxic condition (hWJMSCs-norCL) towards several cancer cell lines including ovarian (SKOV3), cervical (HeLa), tongue squamosa (HSC3), and liver (HepG2) proliferation. Human mesenchymal stem cells (hMSCs), human fibroblast cells, and mouse fibroblast cells (NIH3T3) were used as control. Both hWJMSCs-CL grown in hypoxic and normoxic conditions could inhibit the proliferation of various cancer cell lines with median inhibitory concentration (IC50) 21.09 - 95.92 μg/mL and could cause low inhibition of the normal cells with IC50 409.19 - 799.74 μg/mL. The hWJMSCs-hypoCL and hWJMSC-norCL could inhibit various cancer and were not harmful to normal cells.

scholarly journals Decreased Inhibition of Proliferation and Induction of Apoptosis in Breast Cancer Cell Lines (T47D and MCF7) from Treatment with Conditioned Medium Derived from Hypoxia-Treated Wharton’s Jelly MSCs Compared with Normoxia-Treated MSCs

2021 ◽  
Author(s):  
Wahyu Widowati ◽  
Harry Murti ◽  
Halida Widyastuti ◽  
Dian Ratih Laksmitawati ◽  
Rizal Rizal ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Anticancer Activity ◽  
Cell Lines ◽  
Cytotoxic Effect ◽  
Wharton’S Jelly ◽  
Breast Cancer Cell Lines ◽  
Self Renewal ◽  
Wharton's Jelly

Background: Mesenchymal stem cells (MSCs) are an appealing source of adult stem cells for cell therapy due to the high rate of proliferation, self-renewal capability, and applicable therapy. Wharton’s jelly (WJ), the main component of the umbilical cord extracellular matrix, comprises multipotent stem cells with a high proliferation rate and self-renewal capability and has anti-cancer properties. MSCs have been reported to secrete a variety of cytokines that have a cytotoxic effect in various cancers. Oxygen tension affects MSCs proliferation, cytokines level but no in surface markers expression, MSCs’ differentiation. We explored the cytotoxic effect and inducing apoptosis of Wharton’s jelly derived mesenchymal stem cells (WJMSCs) secretions from normoxic WJMSCs (WJMSCs-norCM) (CM: conditioned medium) and hypoxic WJMSCs (WJMSCs-hypoCM) in breast cancer cell lines (T47D and MCF7). Materials and Methods: Cytotoxic activity was determined using the MTS assay. RT-PCR was performed to measure the expression of apoptosis-inducing genes, specifically P53, BAX, and CASP9, and the antiapoptotic gene BCL-2. Results: WJMSCs-norCM and WJMSCs-hypoCM were potent inhibitors of the proliferation in both cell lines. WJMSCs-norCM had more anticancer activity in T47D and MCF7. The IC50 value of WJMSCs-norCM on MCF7 was 42.34%, and on T47D was 42.36%. WJMSCs-norCM significantly induced the gene expression of apoptotic P53, BAX, and CASP9 and insignificantly decreased the antiapoptotic gene BCL-2 in both MCF7 and T47D cells. WJMSCs-CM has anticancer activity by inducing P53, BAX, and CASP9 apoptotic genes. Conclusion: WJMSCs-norCM has more anticancer activity than WJMSCs-hypoCM.

scholarly journals Immunomodulatory Properties of Wharton’s Jelly-Derived Mesenchymal Stem Cells from Three Anatomical Segments of Umbilical Cord

2021 ◽  
Vol 50 (6) ◽  
pp. 1715-1726
Author(s):  
Jezamine Lim ◽  
Sue Ping Eng ◽  
Wei Yen Yeoh ◽  
Yik Wan Low ◽  
Nur Mohd Shafwan bin Jusoh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Wharton’S Jelly ◽  
Wharton's Jelly ◽  
Cell Based Therapy ◽  
Hla Dr ◽  
Significant Difference

Mesenchymal stem cells (MSCs) are multipotent progenitor cells that are reported to be immune-privileged and immune-evasive. MSCs are capable of differentiating into specific cell types for subsequent use in cell-based therapy. They express low levels of human leucocyte antigen (HLA)-ABC and no HLA-DR. Wharton’s jelly-derived MSCs (WJ-MSCs) were also found to express human leukocyte antigen G (HLA-G), which renders them immunosuppressive. This study aimed to determine whether cultured WJ-MSCs retain their immune-privileged and immune-evasive properties after cell differentiation, and whether these properties differ among MSCs derived from different anatomical segments of the umbilical cord. Umbilical cords of healthy pregnant mothers undergoing caesarean section were obtained and grouped by three anatomical segments: fetal, middle, and maternal segments. WJ-MSCs were isolated, culture-expanded, and differentiated into osteogenic cells. Expression of HLA-DR, HLA-ABC, and HLA-G were quantified using flow cytometry. Both undifferentiated and osteodifferentiated WJ-MSCs were subsequently co-cultured with allogeneic peripheral blood mononuclear cells with/without lipopolysaccharide (LPS) stimulation for five days. Lymphocyte proliferation assay was performed using carboxyfluorescein succinimidyl ester (CFSE) as a tracker. Our results showed no significant difference existed in the HLA profiles among WJ-MSCs from different segments and between WJ-MSCs with and without osteogenic differentiation. Mean levels for HLA-G, HLABC, and HLA-DR were 24.82±17.64, 52.50±18.41, and 1.00±1.68%, respectively. Stimulation with LPS and WJ-MSCs increased peripheral blooc mononuclear cells (PBMC) proliferation. However, PBMC proliferation was significantly lower when PBMCs were co-cultured with osteodifferentiated WJ-MSCs (p < .05; with LPS stimulation and p < .001 without LPS stimulation) than when they were co-cultured with undifferentiated WJ-MSCs. These findings suggest that cultured WJ-MSCs stimulate lymphocyte proliferation and are not immune-privileged. Osteodifferentiated WJ-MSCs reduced the immunogenicity of WJ-MSCs, and this reduction in PBMC proliferation was even more pronounced in the presence of LPS (p < .05). In conclusion, cultured WJ-MSCs are not immune-privileged. Osteodifferentiated WJ-MSCs are less immunogenic than undifferentiated WJ-MSCs, in which case hypoimmunogenicity is more profound under LPS-stimulated conditions.

scholarly journals Assessment of the Neuroprotective and Stemness Properties of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells under Variable (5% vs. 21%) Aerobic Conditions

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 717
Author(s):  
Ewelina Tomecka ◽  
Wioletta Lech ◽  
Marzena Zychowicz ◽  
Anna Sarnowska ◽  
Magdalena Murzyn ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Atmospheric Oxygen ◽  
Neuroprotective Effect ◽  
Culture Conditions ◽  
Wharton’S Jelly ◽  
Oxygen Level ◽  
Wharton's Jelly ◽  
Cell Based Therapy ◽  
Oxygen Conditions

To optimise the culture conditions for human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) intended for clinical use, we investigated ten different properties of these cells cultured under 21% (atmospheric) and 5% (physiological normoxia) oxygen concentrations. The obtained results indicate that 5% O2 has beneficial effects on the proliferation rate, clonogenicity, and slowdown of senescence of hWJ-MSCs; however, the oxygen level did not have an influence on the cell morphology, immunophenotype, or neuroprotective effect of the hWJ-MSCs. Nonetheless, the potential to differentiate into adipocytes, osteocytes, and chondrocytes was comparable under both oxygen conditions. However, spontaneous differentiation of hWJ-MSCs into neuronal lineages was observed and enhanced under atmospheric oxygen conditions. The cells relied more on mitochondrial respiration than glycolysis, regardless of the oxygen conditions. Based on these results, we can conclude that hWJ-MSCs could be effectively cultured and prepared under both oxygen conditions for cell-based therapy. However, the 5% oxygen level seemed to create a more balanced and appropriate environment for hWJ-MSCs.

scholarly journals Enhanced Hepatogenic Differentiation of Human Wharton’s Jelly–Derived Mesenchymal Stem Cells by Using Three-Step Protocol

2019 ◽  
Vol 20 (12) ◽  
pp. 3016 ◽  
Author(s):  
Wachira Panta ◽  
Sumeth Imsoonthornruksa ◽  
Ton Yoisungnern ◽  
Sanong Suksaweang ◽  
Mariena Ketudat-Cairns ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Glycogen Storage ◽  
Wharton’S Jelly ◽  
Wharton's Jelly ◽  
Lineage Differentiation ◽  
Cell Based Therapy ◽  
Hepatogenic Differentiation ◽  
Pre Treatment

Currently, human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) are an attractive source of stem cells for cell-based therapy, owing to their ability to undergo self-renewal and differentiate into all mesodermal, some neuroectodermal, and endodermal progenies, including hepatocytes. Herein, this study aimed to investigate the effects of sodium butyrate (NaBu), an epigenetic regulator that directly inhibits histone deacetylase, on hepatic endodermal lineage differentiation of hWJ-MSCs. NaBu, at 1 mM, optimally promoted endodermal differentiation of hWJ-MSCs, along with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) supplementation (EGF + bFGF + 1 mM NaBu). CXCR4, HNF3β, SOX17 (endodermal), and GATA6 (mesendodermal) mRNAs were also up-regulated (p < 0.001). Immunocytochemistry and a Western blot analysis of SOX17 and HNF3β confirmed that the EGF + bFGF + 1 mM NaBu condition was appropriately pre-treated with hWJ-MSCs before hepatogenic differentiation. Furthermore, the hepatogenic medium + NaBu pre-treatment up-regulated hepatoblast (AFP and HNF3β) and hepatic (CK18 and ALB) markers, and increased the proportion of mature hepatocyte functions, including G6P, C/EBPα, and CYP2B6 mRNAs, glycogen storage and urea secretion. The hepatogenic medium + NaBu in the pre-treatment step can induce hWJ-MSC differentiation toward endodermal, hepatoblastic, and hepatic lineages. Therefore, the hepatogenic medium + NaBu pre-treatment for differentiating hWJ-MSCs could represent an alternative protocol for cell-based therapy and drug screening in clinical applications.

scholarly journals Regenerative Potential of the Product “CardioCell” Derived from the Wharton’s Jelly Mesenchymal Stem Cells for Treating Hindlimb Ischemia

2019 ◽  
Vol 20 (18) ◽  
pp. 4632 ◽  
Author(s):  
Musiał-Wysocka ◽  
Kot ◽  
Sułkowski ◽  
Majka
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Wharton’S Jelly ◽  
Great Promise ◽  
Therapeutic Effects ◽  
Hindlimb Ischemia ◽  
Wharton's Jelly ◽  
Cell Based Therapy ◽  
Therapeutic Benefits

In recent years, mesenchymal stem cells (MSCs) have emerged as a promising therapeutic modality in regenerative medicine. They hold great promise for treating civilization-wide diseases, including cardiovascular diseases, such as acute myocardial infarction and critical limb ischemia. MSCs isolated from Wharton’s jelly (WJ-MSCs) may be utilized in both cell-based therapy and vascular graft engineering to restore vascular function, thereby providing therapeutic benefits for patients. The efficacy of WJ-MSCs lies in their multipotent differentiation ability toward vascular smooth muscle cells, endothelial cells and other cell types, as well as their capacity to secrete various trophic factors, which are potent in promoting angiogenesis, inhibiting apoptosis and modulating immunoreaction. Ischemic limb disease is caused by insufficient nutrient and oxygen supplies resulting from damaged peripheral arteries. The lack of nutrients and oxygen causes severe tissue damage in the limb, thereby resulting in severe morbidities and mortality. The therapeutic effects of the conventional treatments are still not sufficient. Cell transplantations in small animal model (mice) are vital for deciphering the mechanisms of MSCs’ action in muscle regeneration. The stimulation of angiogenesis is a promising strategy for the treatment of ischemic limbs, restoring blood supply for the ischemic region. In the present study, we focus on the therapeutic properties of the human WJ-MSCs derived product, Cardio. We investigated the role of CardioCell in promoting angiogenesis and relieving hindlimb ischemia. Our results confirm the healing effect of CardioCell and strongly support the use of the WJ-MSCs in regenerative medicine.

scholarly journals Comparative Evaluation of Human Mesenchymal Stem Cells of Fetal (Wharton's Jelly) and Adult (Adipose Tissue) Origin during ProlongedIn VitroExpansion: Considerations for Cytotherapy

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
I. Christodoulou ◽  
F. N. Kolisis ◽  
D. Papaevangeliou ◽  
V. Zoumpourlis
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
High Efficiency ◽  
Early Stage ◽  
Wharton’S Jelly ◽  
Phenotypic Data ◽  
Biological Origin ◽  
Wharton's Jelly

Mesenchymal stem cells (MSCs) are somatic cells with a dual capacity for self-renewal and differentiation, and diverse therapeutic applicability, both experimentally and in the clinic. These cells can be isolated from various human tissues that may differ anatomically or developmentally with relative ease. Heterogeneity due to biological origin orin vitromanipulation is, nevertheless, considerable and may equate to differences in qualitative and quantitative characteristics which can prove crucial for successful therapeutic use. With this in mind, in the present study we have evaluated the proliferation kinetics and phenotypic characteristics of MSCs derived from two abundant sources, that is, fetal umbilical cord matrix (Wharton's jelly) and adult adipose tissue (termed WJSC and ADSC, resp.) during prolongedin vitroexpansion, a process necessary for obtaining cell numbers sufficient for clinical application. Our results show that WJSC are derived with relatively high efficiency and bear a substantially increased proliferation capacity whilst largely sustaining the expression of typical immunophenotypic markers, whereas ADSC exhibit a reduced proliferation potential showing typical signs of senescence at an early stage. By combining kinetic with phenotypic data we identify culture thresholds up to which both cell types maintain their stem properties, and we discuss the practical implications of their differences.

Sign in / Sign up

Export Citation Format

Share Document