A Modified MTS Proliferation Assay for Suspended Cells to Avoid the Interference by Hydralazine and β-Mercaptoethanol

Background: Laminin, a member of the Extracellular Matrix (ECM), is a glycoprotein that is used as a factor that affects cell adhesion, proliferation, survival, and differentiation. Of these, five globular domains (LG domains) of the alpha chain play an important role in influencing the cell by binding to the integrin. Objective: This study aimed to evaluate the ability of globular domains 1-3 of laminin alpha2 (rhLAMA2LG1-3) in maintaining the pluripotency of human Mesenchymal Stem Cells (hMSCs), which are widely used in regenerative medicine. Methods: hMSCs were grown in the medium supplemented with rhLAMA2LG1-3, then the effect of the protein on hMSCs were confirmed through cell adhesion assay, proliferation assay and RTPCR. Results: rhLAMA2LG1-3 expressed in Escherichia coli has a molecular weight of 70 kDa, at 1 µg/ml concentration of rhLAMA2LG1-3, the attachment and proliferation of hMSCs were approximately 3.18-fold and 1.67-fold, respectively, more efficient than those of untreated controls. In addition, the undifferentiated state and degree of stemness of hMSCs were measured, on the basis of CD90 and CD105 levels. In the rhLAMA2LG1-3-treated hMSCs, the expression levels of CD90 and CD105 increased by 2.83-fold and 1.62-fold, respectively, compared to those in untreated controls. Conclusion: rhLAMA2LG1-3 can be potentially used in stem cell therapy to improve the viability and maintain the undifferentiated state of hMSCs.

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Mechanical oscillations enhance gene delivery into suspended cells

Scientific Reports ◽

10.1038/srep22824 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 4

Author(s):

Z. L. Zhou ◽

X. X. Sun ◽

J. Ma ◽

C. H. Man ◽

A. S. T. Wong ◽

...

Keyword(s):

Gene Delivery ◽

Mechanical Oscillations ◽

Suspended Cells

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Analysis of Estrogenic Activity in Maryland Coastal Bays Using the MCF-7 Cell Proliferation Assay

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18126254 ◽

2021 ◽

Vol 18 (12) ◽

pp. 6254

Author(s):

Rehab Elfadul ◽

Roman Jesien ◽

Ahmed Elnabawi ◽

Paulinus Chigbu ◽

Ali Ishaque

Keyword(s):

Cell Proliferation ◽

Estrogenic Activity ◽

Landing Site ◽

Negative Control ◽

Proliferation Assay ◽

Negative Effects ◽

Cell Proliferation Assay ◽

Coastal Bays ◽

Maryland Coastal Bays ◽

Mcf 7

Contaminants of Emerging Concern (CECs) with estrogenic or estrogenic-like activity have been increasingly detected in aquatic environments and have been an issue of global concern due to their potential negative effects on wildlife and human health. This study used the MCF-7 cell proliferation assay (E-Screen) to assess the estrogenic activity profiles in Maryland Coastal Bays (MCBs), a eutrophic system of estuaries impacted by human activities. Estrogenic activity was observed in all study sites tested. Water samples from MCBs increased MCF-7 cell proliferation above the negative control from 2.1-fold at site 8, located in Sinepuxent Bay close to the Ocean City Inlet, to 6.3-fold at site 6, located in Newport Bay. The proliferative effects of the sediment samples over the negative control ranged from 1.9-fold at the Assateague Island National Seashore site to 7.7-fold at the Public Landing site. Moreover, elevated cell proliferation (p < 0.05) was observed when cells were co-exposed with 17ß-Estradiol (E2), while reduction in cell proliferation was observed when cells were co-exposed with the antagonist ICI 182, 780 suggesting that cell proliferative effects were primarily mediated by the estrogen receptor (ER). These results suggest the occurrence of some estrogenic or hormonal-like compounds in the MCBs and are consistent with our previous findings based on vitellogenin analyses.

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Rapid Evaluation ofAntrodia camphorataNatural Products and Derivatives in Tumourigenic Liver Progenitor Cells with a Novel Cell Proliferation Assay

ChemMedChem ◽

10.1002/cmdc.200900238 ◽

2009 ◽

Vol 4 (10) ◽

pp. 1657-1667 ◽

Cited By ~ 13

Author(s):

Scott G. Stewart ◽

Louisa A. Ho ◽

Marta E. Polomska ◽

Alexander T. Percival ◽

George C. T. Yeoh

Keyword(s):

Cell Proliferation ◽

Progenitor Cells ◽

Rapid Evaluation ◽

Proliferation Assay ◽

Cell Proliferation Assay

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Proliferation assay of mouse embryonic stem (ES) cells exposed to atmospheric-pressure plasmas at room temperature

Journal of Physics D Applied Physics ◽

10.1088/0022-3727/47/44/445402 ◽

2014 ◽

Vol 47 (44) ◽

pp. 445402 ◽

Cited By ~ 7

Author(s):

Taichi Miura ◽

Ayumi Ando ◽

Kazumi Hirano ◽

Chika Ogura ◽

Tatsuya Kanazawa ◽

...

Keyword(s):

Atmospheric Pressure ◽

Room Temperature ◽

Es Cells ◽

Embryonic Stem ◽

Proliferation Assay ◽

Atmospheric Pressure Plasmas

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Thiol-sensitive sites in cell adhesion. Decreased entry of SH-binding reagents into attached BHK cells

Biochemical Journal ◽

10.1042/bj2080473 ◽

1982 ◽

Vol 208 (2) ◽

pp. 473-478 ◽

Cited By ~ 7

Author(s):

D D McAbee ◽

F Grinnell

Keyword(s):

Cell Adhesion ◽

Small Molecules ◽

Cell Spreading ◽

The Other ◽

Bhk Cells ◽

Sh Groups ◽

Nitrobenzoic Acid ◽

Cytoplasmic Proteins ◽

Adhesion Process ◽

Suspended Cells

Studies were carried out to learn more about the critical SH groups involved in cell spreading. Pretreatment of suspended baby hamster kidney (BHK) cells with 3 mM-iodoacetate or iodoacetamide for 10 min at 4 degrees C completely inhibited the ability of the cells to spread on fibronectin-coated substrata. If, however, BHK cells were permitted to attach and spread before being treated with the SH-binding reagents, and then harvested by trypsinization and assayed for spreading on fibronectin-coated substrata, there was no inhibition of cell spreading. The extent of prior attachment required before the cells became insensitive to the SH-binding reagents was tested and was found to occur early during the cell adhesion process, before any cell spreading was observed. In analytical experiments, there did not appear to be any difference in the total number of SH groups between suspended or spread cells as determined with 5,5′-dithiobis-(2-nitrobenzoic acid). The uptake of radiolabelled iodoacetate into intact spread cells, however, was found to be 3.5 times less than that found with suspended cells. On the other hand, the distribution of incorporated radioactivity into suspended and spread cells was similar. Most of the radioactivity (approximately 70%) was incorporated into small molecules (e.g. glutathione and cysteine), less (approximately 20%) was incorporated into cytoplasmic proteins, and the least incorporation (approximately 10%) was into the cell cytoskeleton. The data are interpreted to indicate there is a decreased permeability of spread cells to the SH-binding reagents.

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