A new clinical-scale serum-free xeno-free medium efficient in ex vivo amplification of mesenchymal stromal cells does not support mesenchymal stem cells

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.

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Neural differentiation of canine mesenchymal stem cells/multipotent mesenchymal stromal cells

BMC Veterinary Research ◽

10.1186/s12917-020-02493-2 ◽

2020 ◽

Vol 16 (1) ◽

Author(s):

Sonja Prpar Mihevc ◽

Vesna Kokondoska Grgich ◽

Andreja Nataša Kopitar ◽

Luka Mohorič ◽

Gregor Majdič

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Neural Differentiation ◽

Multipotent Mesenchymal Stromal Cells

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Human Mesenchymal Stem Cells Display Reduced Expression of CD105 after Culture in Serum-Free Medium

Stem Cells International ◽

10.1155/2013/698076 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 46

Author(s):

Peter Mark ◽

Mandy Kleinsorge ◽

Ralf Gaebel ◽

Cornelia A. Lux ◽

Anita Toelk ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Ex Vivo ◽

Human Mesenchymal Stem Cells ◽

Cell Types ◽

Growth Media ◽

Clinical Use ◽

Differentiation Potential ◽

Serum Free ◽

The Impact

Human Mesenchymal Stem Cells (hMSCs) present a promising tool for regenerative medicine. However,ex vivoexpansion is necessary to obtain sufficient cells for clinical therapy. Conventional growth media usually contain the critical component fetal bovine serum. For clinical use, chemically defined media will be required. In this study, the capability of two commercial, chemically defined, serum-free hMSC growth media (MSCGM-CD and PowerStem) for hMSC proliferation was examined and compared to serum-containing medium (MSCGM). Immunophenotyping of hMSCs was performed using flow cytometry, and they were tested for their ability to differentiate into a variety of cell types. Although the morphology of hMSCs cultured in the different media differed, immunophenotyping displayed similar marker patterns (high expression of CD29, CD44, CD73, and CD90 cell surface markers and absence of CD45). Interestingly, the expression of CD105 was significantly lower for hMSCs cultured in MSCGM-CD compared to MSCGM. Both groups maintained mesenchymal multilineage differentiation potential. In conclusion, the serum-free growth medium is suitable for hMSC culture and comparable to its serum-containing counterpart. As the expression of CD105 has been shown to positively influence hMSC cardiac regenerative potential, the impact of CD105 expression onto clinical use after expansion in MSCGM-CD will have to be tested.

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Apoptosis, DAP-Kinase1 Expression and the Influences of Cytokine Milieu and Mesenchymal Stromal Cells on Ex Vivo Expansion of Umbilical Cord Blood-Derived Hematopoietic Stem Cells

Indian Journal of Hematology and Blood Transfusion ◽

10.1007/s12288-015-0545-y ◽

2015 ◽

Vol 32 (1) ◽

pp. 67-77 ◽

Cited By ~ 2

Author(s):

Naser Amirizadeh ◽

Arezoo Oodi ◽

Roya Mehrasa ◽

Mahin Nikougoftar

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Cord Blood ◽

Umbilical Cord ◽

Mesenchymal Stromal Cells ◽

Umbilical Cord Blood ◽

Stromal Cells ◽

Ex Vivo ◽

Hematopoietic Stem ◽

Cytokine Milieu

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Comparative Study of the Biological Characteristics of Serum-Free and Fetal Bovine Serum-Contained Medium Cultured Umbilical Cord-Derived Mesenchymal Stem Cells

Blood ◽

10.1182/blood.v120.21.4737.4737 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4737-4737 ◽

Cited By ~ 1

Author(s):

Guanghua Chen ◽

Ting Yang ◽

Man Qiao ◽

Huiwen Liu ◽

Wu Depei

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

T Cell ◽

Umbilical Cord ◽

Complete Medium ◽

Human Umbilical Cord ◽

Serum Free Medium ◽

Free Medium ◽

Early Passage ◽

Serum Free

Abstract Abstract 4737 Objective: To compare the difference of biological characteristics between human umbilical cord-derived mesenchymal stem cells (UC-MSC) cultured by serum free medium and fetal bovine serum-contained complete medium and to create a xenogeneic protein-free UC-MSC culture system. Methods: Healthy human umbilical cord segments were digested with collagenase. Umbilical cord-derived mesenchymal stem cells were cultured by serum free MesenCult-XF medium and FBS-based αMEM complete medium. We analysed the morphology, immunophenotype, expansion potential, trilineage differentiation potential, karyotype and immunosuppression of early passage of UC-MSC. Results: The average cell diameters of UC-MSC in suspension cultured by serum free medium and FBS-based medium are 26 (18–39) μm and 35 (20–61) μm, respectively. Cell expansion folds with serum free medium and FBS-based medium were (5.2±0.2) and (3.5±0.1) in the first five passage, respectively. The expansion potential of MSCs was significantly higher with serum free medium compared to FBS-based medium (P<0.05). A panel of markers as CD29, CD44, CD90, CD73, CD105 and HLA-ABC were expressed by human UC-MSC. Hematopoietic lineage markers CD34, CD45 and HLA-DR were not detectable on UC-MSC. The cpm were (4.57±0.14)×104, (2.04±0.16)×104 and(0.42±0.04)×104 when serum free medium cultured MSCs were added to the cultures at ratios MSCs/T cell of 1:100, 1:10 and 1:5. While the cpm were (4.57±0.14)×104, (2.04±0.16)×104 and(0.42±0.04)×104when serum free medium cultured UC-MSCs were added to the cultures. The immunosuppressive potential of serum free medium-cultured UC-MSC was higher than that of serum-contained medium cultured UC-MSC at three different ratios MSC/T cell (P<0.05). Conclusion Compare with serum-contained medium cultured early passage of UC-MSC, the cell diameter of serum free medium cultured MSCs was smaller and the expansion potential was higher. No xenogeneic proteins were presented in UC-MSC preparation when UC-MSC was cultured with serum free medium. Human UC-MSC suppresses T-cell proliferation in a dose-dependent manner. The immunosuppressive potential of UC-MSC was higher when cultured in serum free medium compared with FBS-based medium. Disclosures: No relevant conflicts of interest to declare.

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