scholarly journals Generation of Therapeutically Potent Spheroids from Human Endometrial Mesenchymal Stem/Stromal Cells

2021 ◽  
Vol 11 (6) ◽  
pp. 466
Author(s):  
Alisa Domnina ◽  
Larisa Alekseenko ◽  
Irina Kozhukharova ◽  
Olga Lyublinskaya ◽  
Mariia Shorokhova ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Tissue Repair ◽  
Monolayer Culture ◽  
Great Promise ◽  
Narrow Size Distribution ◽  
Multilineage Differentiation ◽  
Spheroid Formation ◽  
Serum Free ◽  
Multicellular Aggregates ◽  
Free Media

Endometrial mesenchymal stem/stromal cells (eMSCs) hold great promise in bioengineering and regenerative medicine due to their high expansion potential, unique immunosuppressive properties and multilineage differentiation capacity. Usually, eMSCs are maintained and applied as a monolayer culture. Recently, using animal models with endometrial and skin defects, we showed that formation of multicellular aggregates known as spheroids from eMSCs enhances their tissue repair capabilities. In this work, we refined a method of spheroid formation, which makes it possible to obtain well-formed aggregates with a narrow size distribution both at early eMSC passages and after prolonged cultivation. The use of serum-free media allows this method to be used for the production of spheroids for clinical purposes. Wound healing experiments on animals confirmed the high therapeutic potency of the produced eMSC spheroids in comparison to the monolayer eMSC culture.

scholarly journals Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Samatha Bhat ◽  
Pachaiyappan Viswanathan ◽  
Shashank Chandanala ◽  
S. Jyothi Prasanna ◽  
Raviraja N. Seetharam
Keyword(s):  
Bone Marrow ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Population Doubling ◽  
Seeding Density ◽  
Population Doubling Time ◽  
Differentiation Potential ◽  
Serum Free ◽  
Safety Issues ◽  
Free Media

AbstractBone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. Although therapeutic value of MSCs is now being established through many clinical trials, issues have been raised regarding their expansion as per regulatory guidelines. Fetal bovine serum usage in cell therapy poses difficulties due to its less-defined, highly variable composition and safety issues. Hence, there is a need for transition from serum-based to serum-free media (SFM). Since SFM are cell type-specific, a precise analysis of the properties of MSCs cultured in SFM is required to determine the most suitable one. Six different commercially available low serum/SFM with two different seeding densities were evaluated to explore their ability to support the growth and expansion of BM-MSCs and assess the characteristics of BM-MSCs cultured in these media. Except for one of the SFM, all other media tested supported the growth of BM-MSCs at a low seeding density. No significant differences were observed in the expression of MSC specific markers among the various media tested. In contrary, the population doubling time, cell yield, potency, colony-forming ability, differentiation potential, and immunosuppressive properties of MSCs varied with one another. We show that SFM tested supports the growth and expansion of BM-MSCs even at low seeding density and may serve as possible replacement for animal-derived serum.

scholarly journals Human bone marrow-derived mesenchymal stromal cells cultured in serum-free media demonstrate enhanced antifibrotic abilities via prolonged survival and robust regulatory T cell induction in murine bleomycin-induced pulmonary fibrosis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Shun Takao ◽  
Taku Nakashima ◽  
Takeshi Masuda ◽  
Masashi Namba ◽  
Shinjiro Sakamoto ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Pulmonary Fibrosis ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Pulmonary Inflammation ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Distributed Data ◽  
Serum Free ◽  
Free Media

Abstract Background Mesenchymal stromal cells (MSCs) are a potential therapeutic tool for pulmonary fibrosis. However, ex vivo MSC expansion using serum poses risks of harmful immune responses or unknown pathogen infections in the recipients. Therefore, MSCs cultured in serum-free media (SF-MSCs) are ideal for clinical settings; however, their efficacy in pulmonary fibrosis is unknown. Here, we investigated the effects of SF-MSCs on bleomycin-induced pulmonary inflammation and fibrosis compared to those of MSCs cultured in serum-containing media (S-MSCs). Methods SF-MSCs and S-MSCs were characterized in vitro using RNA sequence analysis. The in vivo kinetics and efficacy of SF-MSC therapy were investigated using a murine model of bleomycin-induced pulmonary fibrosis. For normally distributed data, Student’s t test and one-way repeated measures analysis of variance followed by post hoc Tukey’s test were used for comparison between two groups and multiple groups, respectively. For non-normally distributed data, Kruskal–Wallis and Mann–Whitney U tests were used for comparison between groups, using e Bonferroni’s correction for multiple comparisons. All tests were two-sided, and P < 0.05 was considered statistically significant. Results Serum-free media promoted human bone marrow-derived MSC expansion and improved lung engraftment of intravenously administered MSCs in recipient mice. SF-MSCs inhibited the reduction in serum transforming growth factor-β1 and the increase of interleukin-6 in both the serum and the bronchoalveolar lavage fluid during bleomycin-induced pulmonary fibrosis. SF-MSC administration increased the numbers of regulatory T cells (Tregs) in the blood and lungs more strongly than in S-MSC administration. Furthermore, SF-MSCs demonstrated enhanced antifibrotic effects on bleomycin-induced pulmonary fibrosis, which were diminished by antibody-mediated Treg depletion. Conclusions SF-MSCs significantly suppressed BLM-induced pulmonary inflammation and fibrosis through enhanced induction of Tregs into the lungs and corrected the dysregulated cytokine balance. Therefore, SF-MSCs could be a useful tool for preventing pulmonary fibrosis progression without the demerits of serum use.

scholarly journals Efficient differentiation of beige adipocytes from adult human adipose-derived stem/stromal cells

2020 ◽  
Author(s):  
Amar M. Singh ◽  
Liang Zhang ◽  
John Avery ◽  
Stephen Dalton
Keyword(s):  
Stem Cells ◽  
Cell Therapy ◽  
Drug Screening ◽  
Stromal Cells ◽  
High Efficiency ◽  
Disease Modelling ◽  
Serum Free ◽  
Beige Adipocytes ◽  
Free Media ◽  
Stromal Stem Cells

Abstract Beige adipocytes (also known as brite adipocytes) have significant utility for numerous applications, such as drug screening, cell therapy, and disease modelling. However, a high-efficiency protocol from human adult adipose-derived stem/stromal stem cells (ADSC) has not been described. The protocol described here achieves beige adipocyte purities of >92% in a fully-defined, serum-free media cocktail, which enables these downstream applications. This method provides a significant leap forward over previously described, serum-based protocols that were inconsistent and inefficient.

Evaluating the use of Terumo Quantum® cell expansion system for large scale expansion of mesenchymal stem (stromal) cells in xeno- and serum-free media

Cytotherapy ◽  
2017 ◽  
Vol 19 (5) ◽  
pp. S163-S164
Author(s):  
S. Khan ◽  
M. Salkhordeh ◽  
J. Schafhauser ◽  
F. Hussein ◽  
S. Hodgins ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Cell Expansion ◽  
Large Scale ◽  
Serum Free ◽  
Expansion System ◽  
Scale Expansion ◽  
Free Media

scholarly journals Serum-free media for the production of human mesenchymal stromal cells: a review

2013 ◽  
Vol 46 (6) ◽  
pp. 608-627 ◽  
Author(s):  
S. Gottipamula ◽  
M. S. Muttigi ◽  
U. Kolkundkar ◽  
R. N. Seetharam
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Serum Free ◽  
Human Mesenchymal Stromal Cells ◽  
Free Media

CD34+ Cobblestone Area-Forming Cells Are a Valuable Indicator to Screen the Soluble Inhibitors of Hematopoiesis in the Serum of Aplastic Anemia and MDS Patients.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1439-1439
Author(s):  
Shohei Kikuchi ◽  
Masayoshi Kobune ◽  
Kazuyuki Murase ◽  
Satoshi Iyama ◽  
Tsutomu Sato ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Aplastic Anemia ◽  
Stromal Cells ◽  
Conflicts Of Interest ◽  
Hematopoietic Stem ◽  
Serum Free ◽  
Free Media ◽  
Human Stromal Cells ◽  
Stromal Layer

Abstract Abstract 1439 Poster Board I-462 We have previously shown that cobblestone area (CA) could be observed underneath human stromal cells 2 weeks after coculture in serum free condition. However, it was difficult to evaluate 5 week CA-forming cells (CAFC) which is a surrogate indicator of hematopoietic stem cells because human stromal cells could not survive more than 4 weeks in a serum free media, X-VIVO10. In the present study, we optimized serum free media, combination of cytokines and chemical agents such as stem cell factor (SCF), thrombopoietin (TPO), flt3 ligand (FL) with or without delta like protein 4 (DLL4) and GSK inhibitor. In addition, we added the serum derived from aplastic anemia and MDS patients in this coculture system and evaluated the effect of serum on in vitro hematopoiesis. We found that certain serum free media could maintain the layer of human stromal cells more than 8 weeks. By using this serum free media, bone marrow (BM) CD34+ cells could be cocultured for 8 weeks without disruption of the human stromal layer. Five week CAFC could be observed in all condition of cytokine combination with or without chemical inhibitor. However, because cytoplasmic appearance of cells in some CAs is quite irregular without DLL4 or GSK inhibitor, we conducted immunohistochemical staining by using FITC-labeled anti human CD34, CD11b or CD33 antibodies. Surprisingly, we found that CD34+CAFCs were exclusively observed in the presence of DLL4 or GSK inhibitor. Otherwise, CAs was composed of CD11b+/CD45+ hematopoietic cells. Moreover, NOD/SCID repopulating cells were detected in the coculture containing CD34+CAFCs, suggesting that CD34+CAFCs were a valuable indicator of hematopoietic stem cells (HSCs). Using CD34+CAFCs as a surrogate maker of HSCs, we assessed whether the serum derived from patients of aplastic anemia and MDS could inhibit formation of CD34+CAFC. Notably, some of patient's serum could reduce the number of CD34+CAFCs. These results indicated the possibility that CD34+CAFCs underneath human stromal layer could be useful for screening certain soluble factors to inhibit hematopoiesis in aplastic anemia and MDS. Disclosures No relevant conflicts of interest to declare.

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