Fetal bovine serum-free cryopreservation methods for clinical banking of human adipose-derived stem cells

Park, Seah; Lee, Dong Ryul; Nam, Ji Sun; Ahn, Chul Woo; Kim, Haekwon

doi:10.1016/j.cryobiol.2018.02.008

Cited by 27 publications

(16 citation statements)

References 35 publications

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“…One concern is the use of FBS either as a bioink or as cryoprotectant supplement since it introduces the risk of immunological rejection and the transmission of diseases. The alternative is the use of xeno-free cryoprotectant supplements [51,52]. Moreover, for the implementation of autologous cell therapies, there is still the need for primary cell extraction and expansion, making more challenging the treatment of bone injuries that require immediate attention.…”

Section: Discussionmentioning

confidence: 99%

Effects of Cryopreservation on Cell Metabolic Activity and Function of Biofabricated Structures Laden with Osteoblasts

et al. 2020

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Biofabrication and maturation of bone constructs is a long-term task that requires a high degree of specialization. This specialization falls onto the hierarchy complexity of the bone tissue that limits the transfer of this technology to the clinic. This work studied the effects of the short-term cryopreservation on biofabricated osteoblast-containing structures, with the final aim to make them steadily available in biobanks. The biological responses studied include the osteoblast post-thawing metabolic activity and the recovery of the osteoblastic function of 3D-bioprinted osteoblastic structures and beta tricalcium phosphate (β-TCP) scaffolds infiltrated with osteoblasts encapsulated in a hydrogel. The obtained structures were cryopreserved at −80 °C for 7 days using dimethyl sulfoxide (DMSO) as cryoprotectant additive. After thawing the structures were cultured up to 14 days. The results revealed fundamental biological aspects for the successful cryopreservation of osteoblast constructs. In summary, immature osteoblasts take longer to recover than mature osteoblasts. The pre-cryopreservation culture period had an important effect on the metabolic activity and function maintain, faster recovering normal values when cryopreserved after longer-term culture (7 days). The use of β-TCP scaffolds further improved the osteoblast survival after cryopreservation, resulting in similar levels of alkaline phosphatase activity in comparison with the non-preserved structures. These results contribute to the understanding of the biology of cryopreserved osteoblast constructs, approaching biofabrication to the clinical practice.

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Section: Discussionmentioning

confidence: 99%

Effects of Cryopreservation on Cell Metabolic Activity and Function of Biofabricated Structures Laden with Osteoblasts

et al. 2020

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“…Indeed, like all adult cell types, ASCs also significantly decrease cell growth after a limited number of in vitro passages due to a cellular process termed replicative senescence. Nowadays, standard protocols for ASC seeding are lacking, as evidenced by the wide range of cellular concentrations (cell density/cm 2 ) used for replating [ 20 , 27 , 28 , 29 ].…”

Section: Culture and Expansion Of Ascsmentioning

confidence: 99%

“…To avoid FBS use, the addition to cryopreservation medium of different supplements such as human serum albumin (9%), human serum (90%) or knockout serum replacement (90%) to maintain the ASC recovery rate, viability, and growth kinetics at level similar to the conventional medium with 90% FBS, has been recently reported [ 29 ]. Gene expression, immunophenotype and multi-lineage differentiation ability of ASCs are not influenced by the different used cryopreservation media.…”

Section: Svf/asc Cryopreservationmentioning

confidence: 99%

“…Cryopreserved cells can be subjected to freeze–thaw cycles. According to literature data, two rounds of ASC cryopreservation represent the maximum number [ 29 , 54 ]. In particular, when the cells were subjected to three or four freeze-thaw cycles, the viability, immunophenotype, gene expression and multi-lineage potential have been reported to be similar to fresh cells, even if a decreased replicative capacity with a longer doubling time has been reported [ 29 ].…”

Section: Svf/asc Cryopreservationmentioning

confidence: 99%

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Methods of Isolation, Characterization and Expansion of Human Adipose-Derived Stem Cells (ASCs): An Overview

Palumbo

Lombardi

Siragusa

et al. 2018

IJMS

110

102

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Considering the increasing interest in adipose-derived stem cells (ASCs) in regenerative medicine, optimization of methods aimed at isolation, characterization, expansion and evaluation of differentiation potential is critical to ensure (a) the quality of stem cells also in terms of genetic stability; (b) the reproducibility of beneficial effects; and (c) the safety of their use. Numerous studies have been conducted to understand the mechanisms that regulate ASC proliferation, growth and differentiation, however standard protocols about harvesting and processing techniques are not yet defined. It is also important to note that some steps in the procedures of harvesting and/or processing have been reported to affect recovery and/or the physiology of ASCs. Even considering the great opportunity that the ASCs provide for the identification of novel molecular targets for new or old drugs, the definition of homogeneous preparation methods that ensure adequate quality assurance and control, in accordance with current GMPs (good manufacturing practices), is required. Here, we summarize the literature reports to provide a detailed overview of the methodological issues underlying human ASCs isolation, processing, characterization, expansion, differentiation techniques, recalling at the same time their basilar principles, advantages and limits, in particular focusing on how these procedures could affect the ASC quality, functionality and plasticity.

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“…Indeed, it is well known that this approach is not suitable for clinical use of ASCs or their secretome, due to the risk of transmission of disease as well as xenogeneic immune reactions in the transplanted host [33,86]. In this regard, recently, human serum albumin (HSA), human serum (HS), or knockout serum replacement (KSR) have been suggested as useful replacements of FBS for ASC cryopreservation and clinical banking [87]. It is therefore essential to carry out studies using a human serum or synthetic supplementation, which could also be certified for good manufacturing practice (GMP) use of ASCs or their secretome for wound healing.…”

Section: Discussionmentioning

confidence: 99%

Secretome of Adipose Tissue-Derived Stem Cells (ASCs) as a Novel Trend in Chronic Non-Healing Wounds: An Overview of Experimental In Vitro and In Vivo Studies and Methodological Variables

Lombardi

Palumbo

Augello

et al. 2019

IJMS

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Wound healing is a complex process with a linear development that involves many actors in a multistep timeline commonly divided into four stages: Hemostasis, inflammation, proliferation, and remodeling. Chronic non-healing wounds fail to progress beyond the inflammatory phase, thus precluding the next steps and, ultimately, wound repair. Many intrinsic or extrinsic factors may contribute to such an occurrence, including patient health conditions, age-related diseases, metabolic deficiencies, advanced age, mechanical pressure, and infections. Great interest is being focused on the adipose tissue-derived stem cell’s (ASC) paracrine activity for its potential therapeutic impact on chronic non-healing wounds. In this review, we summarize the results of in vitro and in vivo experimental studies on the pro-wound healing effects of ASC-secretome and/or extracellular vesicles (EVs). To define an overall picture of the available literature data, experimental conditions and applied methodologies are described as well as the in vitro and in vivo models chosen in the reported studies. Even if a comparative analysis of the results obtained by the different groups is challenging due to the large variability of experimental conditions, the available findings are undoubtedly encouraging and fully support the use of cell-free therapies for the treatment of chronic non-healing wounds.

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Fetal bovine serum-free cryopreservation methods for clinical banking of human adipose-derived stem cells

Cited by 27 publications

References 35 publications

Effects of Cryopreservation on Cell Metabolic Activity and Function of Biofabricated Structures Laden with Osteoblasts

Effects of Cryopreservation on Cell Metabolic Activity and Function of Biofabricated Structures Laden with Osteoblasts

Methods of Isolation, Characterization and Expansion of Human Adipose-Derived Stem Cells (ASCs): An Overview

Secretome of Adipose Tissue-Derived Stem Cells (ASCs) as a Novel Trend in Chronic Non-Healing Wounds: An Overview of Experimental In Vitro and In Vivo Studies and Methodological Variables

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