scholarly journals Development of a Biodegradable Microcarrier for the Cultivation of Human Adipose Stem Cells (hASCs) with a Defined Xeno- and Serum-Free Medium

2021 ◽  
Vol 11 (3) ◽  
pp. 925
Author(s):  
Francesco Muoio ◽  
Stefano Panella ◽  
Matias Lindner ◽  
Valentin Jossen ◽  
Yves Harder ◽  
...  

Stirred single-use bioreactors in combination with microcarriers (MCs) have established themselves as a technology that has the potential to meet the demands of current and future cell therapeutic markets. However, most of the published processes have been performed using fetal bovine serum (FBS) containing cell culture medium and non-biocompatible MCs. This approach has two significant drawbacks: firstly, the inevitable potential risks associated with the use of FBS for clinical applications; secondly, non-biocompatible MCs have to be removed from the cell suspension before implantation, requiring a step that causes loss of viable cells and adds further costs and complications. This study aimed to develop a new platform based on a chemically defined xeno- and serum-free cell culture medium and biodegradable MC that can support the growth of human adipose stem cells (hASCs) while still preserving their undifferentiated status. A specific combination of components and manufacturing parameters resulted in a MC prototype, called “BR44”, which delivered the desired functionality. MC BR44 allows the hASCs to stick to its surface and grow in a chemically defined xeno- and serum-free medium (UrSuppe). Although the cells’ expansion rate was not as high as with a commercial non-biodegradable standard MC, those cultured on BR44 maintained a better undifferentiated status in both static and dynamic conditions than those cultured on traditional 2D surfaces.

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 466
Author(s):  
Stefano Panella ◽  
Francesco Muoio ◽  
Valentin Jossen ◽  
Yves Harder ◽  
Regine Eibl-Schindler ◽  
...  

Adipose tissue is an abundant source of stem cells. However, liposuction cannot yield cell quantities sufficient for direct applications in regenerative medicine. Therefore, the development of GMP-compliant ex vivo expansion protocols is required to ensure the production of a “cell drug” that is safe, reproducible, and cost-effective. Thus, we developed our own basal defined xeno- and serum-free cell culture medium (UrSuppe), specifically formulated to grow human adipose stem cells (hASCs). With this medium, we can directly culture the stromal vascular fraction (SVF) cells in defined cell culture conditions to obtain hASCs. Cells proliferate while remaining undifferentiated, as shown by Flow Cytometry (FACS), Quantitative Reverse Transcription PCR (RT-qPCR) assays, and their secretion products. Using the UrSuppe cell culture medium, maximum cell densities between 0.51 and 0.80 × 105 cells/cm2 (=2.55–4.00 × 105 cells/mL) were obtained. As the expansion of hASCs represents only the first step in a cell therapeutic protocol or further basic research studies, we formulated two chemically defined media to differentiate the expanded hASCs in white or beige/brown adipocytes. These new media could help translate research projects into the clinical application of hASCs and study ex vivo the biology in healthy and dysfunctional states of adipocytes and their precursors. Following the cell culture system developers’ practice and obvious reasons related to the formulas’ patentability, the defined media’s composition will not be disclosed in this study.


2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Miaomiao Chen ◽  
Xiaoshuang Zuo ◽  
Qinqin Xu ◽  
Rong Wang ◽  
Suhua Fan ◽  
...  

Nanodiamonds (NDs) have been recognized as emerging carbon-based delivery vehicles due to their biocompatibility. NDs were reported to be nontoxic and suited for biomedical applications in the complete cell culture medium. However, in this study, the cytotoxicity of NDs in serum-free medium was studied and indicated that serum proteins in cell culture medium played significant effect on the toxicity of NDs. Therefore, the interaction mechanism between NDs and a serum protein (human serum albumin, HSA) was first investigated by fluorescence quenching technique and circular dichroism (CD) spectrometry. The results suggest that HSA strongly bonds on the NDs surface to form “protein corona,” which not only prevents the aggregation of NDs and improves its stability but also inhibits the cytotoxicity of NDs. In serum-free medium, NDs exhibited obvious toxicity toward the human lung epithelial cell line (BEAS-2B) and showed concentration-dependent cytotoxicity. In the presence of bovine serum albumin (BSA), which shares structural homology and similar properties with HSA, toxicity of NDs was apparently inhibited. Therefore, the interaction between serum protein and NDs should be considered in the understanding of the biological effects of NDs exposure in biomedical applications.


2007 ◽  
Vol 40 (1-2) ◽  
pp. 98-103 ◽  
Author(s):  
Megha S. Even ◽  
Chad B. Sandusky ◽  
Neal D. Barnard ◽  
Jehangir Mistry ◽  
Madhur K. Sinha

1997 ◽  
Vol 20 (3) ◽  
pp. 115-120 ◽  
Author(s):  
E. Lindenbaum ◽  
Y. Har-Shai ◽  
Y. Ullmann ◽  
A. L. Feitelberg ◽  
M. Tendler ◽  
...  

Burns ◽  
1995 ◽  
Vol 21 (2) ◽  
pp. 110-115 ◽  
Author(s):  
E.S. Lindenbaum ◽  
M. Tendler ◽  
D. Beach

2021 ◽  
Vol 12 (2) ◽  
pp. 25
Author(s):  
Francesco Muoio ◽  
Stefano Panella ◽  
Valentin Jossen ◽  
Matias Lindner ◽  
Yves Harder ◽  
...  

Human adipose stem cells (hASCs) are promising candidates for cell-based therapies, but they need to be efficiently expanded in vitro as they cannot be harvested in sufficient quantities. Recently, dynamic bioreactor systems operated with microcarriers achieved considerable high cell densities. Thus, they are a viable alternative to static planar cultivation systems to obtain high numbers of clinical-grade hASCs. Nevertheless, the production of considerable biomass in a short time must not be achieved to the detriment of the cells’ quality. To facilitate the scalable expansion of hASC, we have developed a new serum- and xeno-free medium (UrSuppe) and a biodegradable microcarrier (BR44). In this study, we investigated whether the culture of hASCs in defined serum-free conditions on microcarriers (3D) or on planar (2D) cell culture vessels may influence the expression of some marker genes linked with the immature degree or the differentiated status of the cells. Furthermore, we investigated whether the biomaterials, which form our biodegradable MCs, may affect cell behavior and differentiation. The results confirmed that the quality and the undifferentiated status of the hASCs are very well preserved when they grow on BR44 MCs in defined serum-free conditions. Indeed, the ASCs showed a gene expression profile more compatible with an undifferentiated status than the same cells grown under standard planar conditions.


2011 ◽  
Vol 27 (3) ◽  
pp. 811-823 ◽  
Author(s):  
Roz Alfred ◽  
Jaret Radford ◽  
Jessica Fan ◽  
Kathryn Boon ◽  
Roman Krawetz ◽  
...  

1995 ◽  
Vol 13 (4) ◽  
pp. 389-392 ◽  
Author(s):  
Michael Zang ◽  
Helmut Trautmann ◽  
Christine Gandor ◽  
Ferruccio Messi ◽  
Fred Asselbergs ◽  
...  

2018 ◽  
Vol 7 (12) ◽  
pp. 893-905 ◽  
Author(s):  
Ken Yoshida ◽  
Ayumu Nakashima ◽  
Shigehiro Doi ◽  
Toshinori Ueno ◽  
Tomoe Okubo ◽  
...  

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