scholarly journals Platelet-rich concentrate in serum free medium enhances osteogenic differentiation of bone marrow-derived human mesenchymal stromal cells

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2347 ◽  
Author(s):  
Shani Samuel ◽  
Raja Elina Ahmad ◽  
Thamil Selvee Ramasamy ◽  
Puvanan Karunanithi ◽  
Sangeetha Vasudevaraj Naveen ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Immunocytochemical Staining ◽  
Growth Media ◽  
Serum Free Medium ◽  
Free Medium ◽  
Alizarin Red ◽  
Serum Free ◽  
Time Points ◽  
Human Mesenchymal Stromal Cells

Previous studies have shown that platelet concentrates used in conjunction with appropriate growth media enhance osteogenic differentiation of human mesenchymal stromal cells (hMSCs). However, their potential in inducing osteogenesis of hMSCs when cultured in serum free medium has not been explored. Furthermore, the resulting osteogenic molecular signatures of the hMSCs have not been compared to standard osteogenic medium. We studied the effect of infrequent supplementation (8-day interval) of 15% non-activated platelet-rich concentrate (PRC) in serum free medium on hMSCs proliferation and differentiation throughout a course of 24 days, and compared the effect with those cultured in a standard osteogenic medium (OM). Cell proliferation was analyzed by alamar blue assay. Gene expression of osteogenic markers (Runx2, Collagen1, Alkaline Phosphatase, Bone morphogenetic protein 2, Osteopontin, Osteocalcin, Osteonectin) were analyzed using Q-PCR. Immunocytochemical staining for osteocalcin, osteopontin and transcription factor Runx2 were done at 8, 16 and 24 days. Biochemical assays for the expression of ALP and osteocalcin were also performed at these time-points. Osteogenic differentiation was further confirmed qualitatively by Alizarin Red S staining that was quantified using cetylpyridinium chloride. Results showed that PRC supplemented in serum free medium enhanced hMSC proliferation, which peaked at day 16. The temporal pattern of gene expression of hMSCs under the influence of PRC was comparable to that of the osteogenic media, but at a greater extent at specific time points. Immunocytochemical staining revealed stronger staining for Runx2 in the PRC-treated group compared to OM, while the staining for Osteocalcin and Osteopontin were comparable in both groups. ALP activity and Osteocalcin/DNA level were higher in the PRC group. Cells in the PRC group had similar level of bone mineralization as those cultured in OM, as reflected by the intensity of Alizarin red stain. Collectively, these results demonstrate a great potential of PRC alone in inducing proliferation of hMSCs without any influence from other lineage-specific growth media. PRC alone has similar capacity to enhance hMSC osteogenic differentiation as a standard OM, without changing the temporal profile of the differentiation process. Thus, PRC could be used as a substitute medium to provide sufficient pool of pre-differentiated hMSCs for potential clinical application in bone regeneration.

Accelerated and safe expansion of human mesenchymal stromal cells in animal serum-free medium for transplantation and regenerative medicine

2007 ◽  
Vol 213 (1) ◽  
pp. 18-26 ◽  
Author(s):  
Claudia Lange ◽  
Figen Cakiroglu ◽  
Andrej-Nikolai Spiess ◽  
Heike Cappallo-Obermann ◽  
Judith Dierlamm ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Human Mesenchymal Stromal Cells

Therapeutic Impact of Human Bone Marrow Stromal Cells Expanded by Animal Serum–Free Medium for Cerebral Infarct in Rats

Neurosurgery ◽  
2011 ◽  
Vol 68 (6) ◽  
pp. 1733-1742 ◽  
Author(s):  
Taku Sugiyama ◽  
Satoshi Kuroda ◽  
Yukari Takeda ◽  
Mitsufumi Nishio ◽  
Masaki Ito ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Marrow Stromal Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Therapeutic Impact

scholarly journals Human olfactory mesenchymal stromal cells co-expressing horizontal basal and ensheathing cell proteins in culture

Biomédica ◽  
2020 ◽  
Vol 40 (1) ◽  
pp. 72-88
Author(s):  
Carlos Ayala-Grosso ◽  
Rosalinda Pieruzzini ◽  
Leslie Vargas-Saturno ◽  
José E. Cardier
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Olfactory Mucosa ◽  
Serum Free Medium ◽  
Free Medium ◽  
Phenotypic Markers ◽  
Serum Free ◽  
Fetal Bovine

Introduction: The olfactory neuro-epithelium has an intrinsic capability of renewal during lifetime provided by the existence of globose and horizontal olfactory precursor cells. Additionally, mesenchymal stromal olfactory cells also support the homeostasis of the olfactory mucosa cell population. Under in vitro culture conditions with Dulbecco modified eagle/F12 medium supplemented with 10% fetal bovine serum, tissue biopsies from upper turbinate have generated an adherent population of cells expressing mainly mesenchymal stromal phenotypic markers. A closer examination of these cells has also found co-expression of olfactory precursors and ensheathing cell phenotypic markers. These results were suggestive of a unique property of olfactory mesenchymal stromal cells as potentially olfactory progenitor cells.Objective: To study whether the expression of these proteins in mesenchymal stromal cells is modulated upon neuronal differentiation.Materials and methods: We observed the phenotype of olfactory stromal cells under DMEM/F12 plus 10% fetal bovine serum in comparison to cells from spheres induced by serum-free medium plus growth factors inducers of neural progenitors.Results: The expression of mesenchymal stromal (CD29+, CD73+, CD90+, CD45-), horizontal basal (ICAM-1/CD54+, p63+, p75NGFr+), and ensheathing progenitor cell (nestin+, GFAP+) proteins was determined in the cultured population by flow cytometry. The determination of Oct 3/4, Sox-2, and Mash-1 transcription factors, as well as the neurotrophins BDNF, NT3, and NT4 by RT-PCR in cells, was indicative of functional heterogeneity of the olfactory mucosa tissue sample. Conclusions: Mesenchymal and olfactory precursor proteins were downregulated by serum-free medium and promoted differentiation of mesenchymal stromal cells into neurons and astroglial cells.

Expression of PRV-1 Decreases Dependency of Cells on Growth Factors for Proliferation.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 655-655
Author(s):  
Vahid Afshar-Kharghan ◽  
Jun Li ◽  
Zakar Mnjoyan
Keyword(s):  
Growth Factors ◽  
Cho Cells ◽  
Growth Media ◽  
Maximal Effect ◽  
Serum Free Medium ◽  
Free Medium ◽  
Transfected Cells ◽  
Serum Free ◽  
Cell Lysate ◽  
Cpg Dinucleotide

Abstract Polycythemia rubra vera-1 (PRV-1) is a GPI-linked protein expressed on surface of neutrophils. Polycythemia vera and essential thrombocythemia were found to be associated with an increase in the level of PRV-1 mRNA; however, the function of PRV-1 remains unknown. We have studied the functional effect of expression of PRV-1, both in heterologous cell line and in neutrophils. cDNA encoding PRV-1 was subcloned from a leukocyte cDNA library and expressed in Chinese hamster ovary cells (CHO). After seeding equal numbers of CHO cells stably transfected with PRV-1 or empty plasmid, cell count was conducted at different time intervals and under different concentration of fetal bovine serum (FBS) in growth media. The numbers of PRV-1 transfected cells were higher than sham-transfected cells at 24, 48, 96 and 144 hrs after seeding, and this difference was enhanced with cells growing in a medium with a reduced concentration of serum with the maximal effect seen in serum-free medium. The percentage of apoptotic cells was similar between PRV-1 and sham-transfected cells. These results are suggestive of that CHO cells expressing PRV-1 proliferate faster than sham-transfected cells. The difference in proliferation rate was more significant at lower concentrations of serum and was the highest in serum-free medium. An immunoblot analysis with anti-phosphotyrosine antibody on the whole cell lysate demonstrated that the presence of PRV-1 alters cell signaling. A 60kDa protein band that disappeared after removal of serum in growth medium of sham-transfected cells continued to be present in the PRV-1 transfected cell lysate regardless of the presence or absence of FBS in the media. In order to understand the function of PRV-1 in neutrophils, we used the fact that in 85% of individuals PRV-1 is expressed only by a subgroup of neutrophils. We separated PRV-1 expressing neutrophils of an individual from those lacking this protein in the same donor, using anti-PRV-1 monoclonal antibody. We then compared the gene expression profile of the two groups of neutrophil using DNA microarray technique. Expression of PRV-1 was associated with several folds increase in expression of growth factors (fracture callus-1, X 14.9 times; Insulin-like 3, X 6.7; neural proliferation differentiation, and control-1 or NPDC1, X 3.4 times) and post-translational modifying enzyme of cell surface protein (N-acetylase N sulfotransferase-1, X 18.9 times). Under physiologic conditions, transcriptional regulation of the PRV-1 gene limits its expression to only a certain percentage of neutrophils. In order to investigate the role of epigenetic factors in regulation of transcription of the PRV-1 gene, we analyzed methylation of CpG dinucleotide in promoter of the PRV-1 gene in the genomic DNA obtained from neutrophils expressing PRV-1 to those that don’t express this protein in the same individual. Expression of PRV-1 was associated with a decrease in methylation of the CpG dinucleotide located 2937 bp before initiation codon (45% methylated in PRV-1 negative neutrophils compared to 30% methylated in PRV positive neutrophils). We are currently studying the methylation pattern of CpG islands inside the PRV-1 gene. Studying the effect of DNA methylation on expression of PRV-1 in physiologic conditions, may shed light to the mechanism of overexpression of PRV-1 in MPD.

scholarly journals Serum-Free Medium and Mesenchymal Stromal Cells Enhance Functionality and Stabilize Integrity of Rat Hepatocyte Spheroids

2013 ◽  
Vol 22 (2) ◽  
pp. 299-308 ◽  
Author(s):  
Ji Bao ◽  
James E. Fisher ◽  
Joseph B. Lillegard ◽  
William Wang ◽  
Bruce Amiot ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Rat Hepatocyte ◽  
Serum Free ◽  
Hepatocyte Spheroids

Potential of 5-azacytidine induction decidual stromal cells from maternal human term placenta towards cardiomyocyte-like cells in serum-free medium

2015 ◽  
Vol 16 (3) ◽  
pp. 477-485 ◽  
Author(s):  
Gecai Chen ◽  
Aihuan Yue ◽  
Zhongbao Ruan ◽  
Yigang Yin ◽  
Ruzhu Wang ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Human Term Placenta ◽  
Serum Free ◽  
Term Placenta

scholarly journals A serum-free medium formulation efficiently supports isolation and propagation of canine adipose-derived mesenchymal stem/stromal cells

PLoS ONE ◽  
2019 ◽  
Vol 14 (2) ◽  
pp. e0210250 ◽  
Author(s):  
Laxminarayana R. Devireddy ◽  
Michael Myers ◽  
Rudell Screven ◽  
Zhuoming Liu ◽  
Lynne Boxer
Keyword(s):  
Stromal Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Medium Formulation
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