scholarly journals Study of Human Amniotic Membrane Mesenchymal Stem Cells Using Gelatin and Alginate as Nontoxic Scaffolds

2019 ◽  
Vol 5 ◽  
pp. 1
Author(s):  
Nike Hendrijantini ◽  
Poedjo Hartono ◽  
Helen Susilowati ◽  
Cindy K. Hartono ◽  
Reni P. Daniati ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Amniotic Membrane ◽  
Colorimetric Assay ◽  
Control Group ◽  
Human Amniotic Membrane ◽  
Average Percentage ◽  
Significant Difference

Perinatal mesenchymal stem cells (MSCs), for example, from amniotic membrane, have advantages over adult sources, such as bone marrow, in terms of ease of availability, cell naivety, tissue stem cell abundance, high capacity of proliferation, and less donor site morbidity, because it does not require invasive procedures. Natural polymer scaffolds, such as gelatin and alginate, are biocompatible. Combination of stem cells from amniotic membrane (hAMSCs) and gelatin or alginate as scaffold can be promising. However, cytotoxicity comparison of gelatin and alginate to hAMSCs has not been widely studied. This study was aimed to compare cytotoxicity of gelatin and alginate on hAMSCs in vitro. Isolation and culture were performed on hAMSCs of the healthy full-term pregnancy. In passage 4, Flow Cytometry CD90, CD105, and CD73 phenotype characterization was done. Colorimetric assay of 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was performed to measure the cytotoxicity. There were three sample groups: (control group) hAMSCs with alpha-minimum essential medium (α-MEM) solution as control; (gelatin group) hAMSCs with gelatin; (alginate group) hAMSCs with alginate. Each group consisted of 12 samples. Flow cytometry of hAMSCs expressed 28.78% CD90, 36.95% CD105, and 44.41% CD73 surface markers. No sample depicted toxicity in either gelatin or alginate group, and this is indicated by the average percentage of living cells in gelatin 97.26% and in alginate 98.43%. No statistically significant difference (ρ=0.057) of cytotoxicity was found between gelatin and alginate to hAMSCs. Gelatin and alginate were nontoxic to hAMSCs in vitro.

scholarly journals Comparative in vitro study of the cytotoxicity of gelatine and alginate to human umbilical cord mesenchymal stem cells

2019 ◽  
Vol 52 (1) ◽  
pp. 36
Author(s):  
Nike Hendrijantini
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Colorimetric Assay ◽  
In Vitro Study ◽  
Human Umbilical Cord ◽  
Significant Difference ◽  
Diphenyl Tetrazolium Bromide

Background: Mesenchymal stem cells (MSCs) and scaffold combination constitute a promising approach currently adopted for tissue engineering. Umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are easily obtained and non-invasive. Gelatine and alginate constitute a biocompatible natural polymer scaffold. At present, a cytotoxicity comparison of gelatine and alginate to hUC-MSCs is not widely conducted Purpose: This study aimed to compare the cytotoxicity of gelatine and alginate in hUC-MSCs in vitro. Methods: Isolation and culture were performed on hUC-MSCs derived from healthy full-term neonates. Flow Cytometry CD90, CD105 and CD73 phenotype characterization was performed in passage 4. 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay was performed to measure the cytotoxicity. The three sample groups were: (T1) hUC-MSCs with α-MEM (alpha-minimum essential medium) solution as control; (T2) hUC-MSCs with gelatine; (T3) hUC-MSCs with alginate Results: Flow cytometry of hUC-MSCs displayed positive CD90, CD105 and CD73 surface markers. Gelatine and alginate had no effect on the viability of hUC-MSCs and no statistically significant difference (p>0.05) of cytotoxicity between gelatine and alginate to hUC-MSCs. Conclusion: Gelatine and alginate proved to be non-toxic to hUC-MSCs in vitro.

scholarly journals An In vitro porcine study of repearing articular cartilage with human amniotic membrane epithelial and mesenchymal stem cells

2012 ◽  
Vol 20 ◽  
pp. S273
Author(s):  
E. Muiños-López ◽  
S.M. Díaz-Prado ◽  
T. Hermida-Gómez ◽  
E. Rendal-Vázquez ◽  
I. Fuentes-Boquete ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Articular Cartilage ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane

scholarly journals Biocompatibility of quantum dots (CdSe/ZnS ) in human amniotic membrane-derived mesenchymal stem cells in vitro

2015 ◽  
Vol 159 (2) ◽  
pp. 227-233 ◽  
Author(s):  
Gongping Wang ◽  
Guangwei Zeng ◽  
Caie Wang ◽  
Huasheng Wang ◽  
Bo Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Quantum Dots ◽  
Mesenchymal Stem Cells ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane

Mesenchymal Stem Cells from Human Amniotic Membrane and Umbilical Cord Can Diminish Immunological Response in an in vitro Allograft Model

2016 ◽  
Vol 82 (3) ◽  
pp. 267-275 ◽  
Author(s):  
Filip A. Dabrowski ◽  
Anna Burdzinska ◽  
Agnieszka Kulesza ◽  
Marcin Chlebus ◽  
Beata Kaleta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Amniotic Membrane ◽  
Immunological Response ◽  
Human Amniotic Membrane

scholarly journals Platelet-Derived Microparticles Increase Expression of hTERT in Umbilical Cord Mesenchymal Stem Cells

2018 ◽  
Vol 5 (4) ◽  
pp. 31 ◽  
Author(s):  
Maryam Samareh Salavati Pour ◽  
Fatemeh Hoseinpour Kasgari ◽  
Alireza Farsinejad ◽  
Ahmad Fatemi ◽  
Roohollah Mirzaee Khalilabadi
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Life Span ◽  
Real Time ◽  
Umbilical Cord ◽  
Control Group ◽  
Surface Markers

Introduction: Mesenchymal stem cells (MSCs) are widely studied due to their self- renewal potential and capacity to differentiate into multiple tissues. However, they have a limited life span of several divisions in vitro, which alters various cellular characteristics and reduces their application. Aim: We evaluated the effect of platelet-derived microparticles on gene expression of hTERT, one of the main factors involved in aging and cell longevity. Materials and methods: Umbilical cord MSCs were used for this study. Cells were characterized by evaluating morphology via inverted microscope and identifying associated surface markers using flow cytometry. Platelet-derived microparticles were prepared by centrifuging platelet bags at varying speeds, and their concen- trations were determined by Bradford assay. At 30% confluency, MSCs were treated with 50 μg/mL of microparticles for five days. Then, RNA was extracted and cDNA was synthesized. Quantitative expression of hTERT was assessed using real-time polymerase chain reaction (PCR). Results: Fibroblast-like cells were isolated from umbilical cord tissue and MSCs were identified by the presence of mesenchymal surface markers via flow cytometry. Real- time PCR showed that gene expression of hTERT increased by more than three times when treated with platelet-derived microparticles, in comparison to expression of the control group. Conclusion: We concluded that platelet-derived microparticles may be a potentially safe and effective method to increase hTERT gene expression in MSCs, ultimately prolonging their life span in vitro. 

scholarly journals Homing of adipose stem cells on the human amniotic membrane as a scaffold: A histological study

2020 ◽  
pp. 21-32
Author(s):  
Hooman Zarei ◽  
Abbasali Karimpour ◽  
Ali Reza Khalatbary ◽  
Fereshteh Talebpour Amiri
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Flow Cytometry ◽  
Amniotic Membrane ◽  
Histological Study ◽  
Adipose Stem Cells ◽  
Human Amniotic Membrane ◽  
Tissue Samples ◽  
Differentiated Cells

Background: The human amniotic membrane (HAM) is a suitable and effective scaffold for cell culture and delivery, and adipose-derived stem cells (ADSCs) are an important source of stem cells for transplantation and chondrogenic differentiation. Objective: To assess the practicability of a cryopreserved HAM as a scaffold in cell proliferation and differentiation in vitro. Materials and Methods: In this experimental study, adipose tissue samples were harvested from the inguinal region of male patients aged 15-30 years. Flow cytometry was used to identify CD31, CD45, CD90, and CD105 markers in adipose stem cells. HAM was harvested from donor placenta after cesarean section, washed, trypsin-based decellularized trypsinized decellularized, and used as a scaffold via three methods: 1) ADSCs were differentiated into chondrocytes on cell culture flasks (monolayer method), and after 14 days of culture, the cells were transferred and cultured on both sides of the HAM; 2) ADSCs were cultured and differentiated directly on both sides of the HAM for 14 days (scaffold-mediated differentiation); and 3) chondrocytes were differentiated with micromass culture for 14 days, transferred on HAM, and tissue slides were histologically analyzed qualitatively. Results: Flow cytometry confirmed the presence of mesenchymal stem cells. Histological findings revealed that the cells adhered and grew well on the stromal layer of HAM. Among the three methods, scaffold-mediated differentiation of ADSCs showed the best results. Conclusion: ADSCs have excellent attachment, viability, and differentiation capacity in the stromal side of HAM. Additionally, the direct culture and differentiation of ADSCs on HAM is more suitable than the culture of differentiated cells on HAM. Key words: Amniotic membrane, Scaffold, Chondrogenesis, Differentiation, Mesenchymal stem cell.

scholarly journals Effects of Adipose-Derived Mesenchymal Stem Cells and Human Amniotic Membrane on Sciatic Nerve Repair in Rats

2021 ◽  
Vol 8 (3) ◽  
Author(s):  
Siyawash Xaki ◽  
Afshin Fathi ◽  
Mehdi Ariana ◽  
Hamid Reza Aghayan ◽  
Babak Arjmand ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Sciatic Nerve ◽  
Nerve Injury ◽  
Amniotic Membrane ◽  
Nerve Fibers ◽  
Sciatic Nerve Injury ◽  
Control Group ◽  
Human Amniotic Membrane

Background: Peripheral nerve injuries remain a great challenge for microsurgery despite the significant progress in recent decades. The current gold standard is autogenous nerve grafting with a success rate as low as 50% in long gaps. Current studies have focused on finding alternative methods for bridging nerve defects. Previous data have demonstrated the role of human amniotic membrane in stimulating neural regeneration. On the other hand, adipose-derived mesenchymal stem cells can differentiate into all three germ layers and could support nerve repair. The purpose of this study was to compare the role of the human amniotic membrane with and without adipose tissue stem cells in sciatic nerve injury with gap in rats. Objectives: We aimed to evaluate the effectiveness of the human amniotic membrane with and without adipose-derived mesenchymal stem cells in sciatic nerve injury with gap in rats. Methods: Twenty-four male Wistar rats in four random groups were used in our study. In the first group, the nerve gap was repaired using the inverse resected nerve segment (Control group), the second group was repaired with a human amniotic membrane (AM group), the third group was repaired with an amnion sheet with seeded adipose-derived mesenchymal stem cells (AM/ADMSCs group), and the last group was not repaired, and both stumps were sutured to muscles. Results: All the animals underwent the procedures and survived without complication. The sciatic function index and hot plate test results were significantly improved in the AM and AM/ADMSCs groups compared to the Control group (as a gold standard of care) (P>0.05). Based on histopathology findings, regenerative nerve fibers were seen in the implanted area of both AM and AM/ADMSCs groups; however, nerve fibers were surrounded by significant fibrosis (scar formation) in the AM/ADMSCs group. The axon count in the Control group was significantly higher than both experimental groups (P < 0.01). Conclusions: Our study showed the role of amniotic membrane in the promotion of nerve regeneration in sciatic nerve injury with a gap, but adding adipose-derived mesenchymal stem cells not only has no extra benefits, but also causes more tissue scar.

scholarly journals Effect of Placental Extract on Omentum Mesenchymal Stem Cells Differentiation in NMRI Mice

2017 ◽  
Vol 7 (1) ◽  
pp. 176
Author(s):  
Maryam Sadat Nezhadfazel ◽  
Kazem Parivar ◽  
Nasim Hayati Roodbari ◽  
Mitra Heydari Nasrabadi
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Flow Cytometry ◽  
Endothelial Cells ◽  
Mesenchymal Stem Cells ◽  
Control Group ◽  
Fat Cell ◽  
Nmri Mice ◽  
Differentiated Cells ◽  
Placental Extract

Omentum mesenchymal stem cells (OMSCs) could be induced to differentiate into cell varieties under certain conditions. We studied differentiation of OMSCs induced by using placenta extract in NMRI mice. Mesenchymal stem cells (MSCs) were isolated from omentum and cultured with mice placenta extract. MSCs, were assessed after three passages by flow cytometry for CD90, CD44, CD73, CD105, CD34 markers and were recognized their ability to differentiate into bone and fat cell lines. Placenta extract dose was determined with IC50 test then OMSCs were cultured in DMEM and 20% placenta extract.The cell cycle was checked. OMSCs were assayed on 21 days after culture and differentiated cells were determined by flow cytometry and again processed for flow cytometry. CD90, CD44, CD73, CD105 markers were not expressed, only CD34 was their marker. OMSCs were morphologically observed. Differentiated cells are similar to the endothelial cells. Therefore, to identify differentiated cells, CD31 and FLK1 expression were measured. This was confirmed by its expression. G1 phase of the cell cycle shows that OMSCs compared to the control group, were in the differentiation phase. The reason for the differentiation of MSCs into endothelial cells was the sign of presence of VEGF factor in the medium too high value of as a VEGF secreting source.

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