Serum-free, xeno-free culture media maintain the proliferation rate and multipotentiality of adipose stem cells in vitro

Lindroos, Bettina; Boucher, Shayne; Chase, Lucas G.; Kuokkanen, Hannu; Huhtala, Heini; Haataja, Riina; Vemuri, Mohan C.; Suuronen, Riitta; Miettinen, Susanna

doi:10.3109/14653240903233081

Cited by 199 publications

(221 citation statements)

References 54 publications

Supporting

Mentioning

205

Contrasting

Order By: Relevance

“…Further, to assess whether AT-MSCs retained their immunophenotype after being cultured in media with UC-dFBS or UF-dFBS compared with FBS for 48 h, AT-MSCs ( n = 3) were analysed for surface markers CD34 (clone: 581), CD45RO (clone: UCHL1), CD54 (clone: HA58), CD73 (clone: AD2) and CD105 (clone: 266). A total of 100 000 cells per sample were analysed, and positive expression was defined as the level of fluorescence greater than 99% of the corresponding unstained cell sample [20]. …”

Section: Methodsmentioning

confidence: 99%

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Kornilov

Puhka

Mannerström

et al. 2018

J of Extracellular Vesicle

156

125

View full text Add to dashboard Cite

Fetal bovine serum (FBS) is the most commonly used supplement in studies involving cell-culture experiments. However, FBS contains large numbers of bovine extracellular vesicles (EVs), which hamper the analyses of secreted EVs from the cell type of preference and, thus, also the downstream analyses. Therefore, a prior elimination of EVs from FBS is crucial. However, the current methods of EV depletion by ultracentrifugation are cumbersome and the commercial alternatives expensive. In this study, our aim was to develop a protocol to completely deplete EVs from FBS, which may have wide applicability in cell-culture applications. We investigated different EV-depleted FBS prepared by our novel ultrafiltration-based protocol, by conventionally used overnight ultracentrifugation, or commercially available depleted FBS, and compared them with regular FBS. All sera were characterized by nanoparticle tracking analysis, electron microscopy, Western blotting and RNA quantification. Next, adipose-tissue mesenchymal stem cells (AT-MSCs) and cancer cells were grown in the media supplemented with the three different EV-depleted FBS and compared with cells grown in regular FBS media to assess the effects on cell proliferation, stress, differentiation and EV production. The novel ultrafiltration-based protocol depleted EVs from FBS clearly more efficiently than ultracentrifugation and commercial methods. Cell proliferation, stress, differentiation and EV production of AT-MSCs and cancer cell lines were similarly maintained in all three EV-depleted FBS media up to 96 h. In summary, our ultrafiltration protocol efficiently depletes EVs, is easy to use and maintains cell growth and metabolism. Since the method is also cost-effective and easy to standardize, it could be used in a wide range of cell-culture applications helping to increase comparability of EV research results between laboratories.

show abstract

Section: Methodsmentioning

confidence: 99%

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Kornilov

Puhka

Mannerström

et al. 2018

J of Extracellular Vesicle

156

125

View full text Add to dashboard Cite

show abstract

“…There is evidence that functional ASCs can be expanded directly from lipoaspirate fluids without the need for collagenase digestion [14]. Similarly, multiple groups routinely use porcine-derived trypsin to passage plastic adherent ASCs, and recent studies have documented the equivalent performance of bacterial-derived or corn-derived trypsin products [15][16][17]. Thus, the removal of enzyme reagents is achievable.…”

Section: Enzyme Qualitymentioning

confidence: 99%

“…An optimal human serum reagent would be depleted of antibodies and complement proteins to reduce the risk of cell damage or adverse events. There is the possibility of removing serum entirely from the culture medium [15]. The Regea Institute has demonstrated the use of a commercially available xenoprotein-free product for ASC expansion [15].…”

Section: Serum Alternativesmentioning

confidence: 99%

“…There is the possibility of removing serum entirely from the culture medium [15]. The Regea Institute has demonstrated the use of a commercially available xenoprotein-free product for ASC expansion [15]. While the proprietary nature of the medium leaves the public with questions about its active ingredients, the deposition of a confidential master file with a regulatory agency (FDA and EMEA) would address this concern.…”

Section: Serum Alternativesmentioning

confidence: 99%

See 1 more Smart Citation

Concise Review: Adipose-Derived Stromal Vascular Fraction Cells and Stem Cells: Let's Not Get Lost in Translation

et al. 2011

View full text Add to dashboard Cite

Subcutaneous fat has emerged as an alternative tissue source for stromal/stem cells in regenerative medicine. Over the past decade, international research efforts have established a wealth of basic science and preclinical evidence regarding the differentiation potential and regenerative properties of both freshly processed, heterogeneous stromal vascular fraction cells and culture expanded, relatively homogeneous adipose-derived stromal/stem cells. The stage has been set for clinicians to translate adipose-derived cells from the bench to the bedside; however, this process will involve “development” steps that fall outside of traditional “hypothesis-driven, mechanism-based” paradigm. This concise review examines the next stages of the development process for therapeutic applications of adipose-derived cells and highlights the current state of the art regarding clinical trials. It is recommended that the experiments addressing these issues be reported comprehensively in the peer-review literature. This transparency will accelerate the standardization and reproducibility of adipose-derived cell therapies with respect to their efficacy and safety.

show abstract

“…The hASCs were isolated as previously described 31. The hASCs were expanded in DMEM/F12 (1:1) (Thermo Fisher Scientific Inc., Carlsbad, CA, USA, https://www.thermofisher.com) supplemented with 5% Pooled Human Platelet Lysate (Stemulate; Cook General Bio‐Technology, Indianapolis, IN, USA, http://cookregenterec.com), 1% antibiotics (100 U/ml penicillin and 0,1 mg/ml streptomycin; Thermo Fisher Scientific Inc.), and 1% l ‐glutamine (Thermo Fisher Scientific Inc, GlutaMAX‐100, Indianapolis, IN, USA).…”

Section: Methodsmentioning

confidence: 99%

Functional Outcome of Human Adipose Stem Cell Injections in Rat Anal Sphincter Acute Injury Model

Kuismanen

Juntunen

Narra

et al. 2018

Stem Cells Translational Medicine

Self Cite

View full text Add to dashboard Cite

Anal incontinence is a devastating condition that significantly reduces the quality of life. Our aim was to evaluate the effect of human adipose stem cell (hASC) injections in a rat model for anal sphincter injury, which is the main cause of anal incontinence in humans. Furthermore, we tested if the efficacy of hASCs could be improved by combining them with polyacrylamide hydrogel carrier, Bulkamid. Human ASCs derived from a female donor were culture expanded in DMEM/F12 supplemented with human platelet lysate. Female virgin Sprague‐Dawley rats were randomized into four groups (n = 14–15/group): hASCs in saline or Bulkamid (3 × 105/60 μl) and saline or Bulkamid without cells. Anorectal manometry (ARM) was performed before anal sphincter injury, at two (n = 58) and at four weeks after (n = 33). Additionally, the anal sphincter tissue was examined by micro‐computed tomography (μCT) and the histological parameters were compared between the groups. The median resting and peak pressure during spontaneous contraction measured by ARM were significantly higher in hASC treatment groups compared with the control groups without hASCs. There was no statistical difference in functional results between the hASC‐carrier groups (saline vs. Bulkamid). No difference was detected in the sphincter muscle continuation between the groups in the histology and μCT analysis. More inflammation was discovered in the group receiving saline with hASC. The hASC injection therapy with both saline and Bulkamid is a promising nonsurgical treatment for acute anal sphincter injury. Traditional histology combined with the 3D μCT image data lends greater confidence in assessing muscle healing and continuity. Stem Cells Translational Medicine 2018;7:295–304

show abstract

Serum-free, xeno-free culture media maintain the proliferation rate and multipotentiality of adipose stem cells in vitro

Cited by 199 publications

References 54 publications

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Efficient ultrafiltration‐based protocol to deplete extracellular vesicles from fetal bovine serum

Concise Review: Adipose-Derived Stromal Vascular Fraction Cells and Stem Cells: Let's Not Get Lost in Translation

Functional Outcome of Human Adipose Stem Cell Injections in Rat Anal Sphincter Acute Injury Model

Contact Info

Product

Resources

About