Testing of nine different xeno-free culture media for human embryonic stem cell cultures

Rajala, Kristiina; Hakala, Heidi; Panula, Sarita; Aivio, Suvi; Pihlajamäki, Harri; Suuronen, Riitta; Hovatta, Outi; Skottman, Heli

doi:10.1093/humrep/del523

Cited by 126 publications

(70 citation statements)

References 25 publications

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“…Several attempts to improve hPSCs culture conditions have been reported. These advances include: the derivation of clinical grade hESC and iPSC lines, the use of conditioned media together with Matrigel TM as an attachment substrate for hPSCs culture and the derivation and propagation of hESC lines on human feeder layers in xenofree culture media (Amit et al, 2004;Hovatta et al, 2003;Rajala et al, 2007;Rajala et al, 2010;Richards et al, 2002;Richards et al, 2003;Skottman et al, 2006;Unger et al, 2008). S o m e a p p r o a c h e s h a v e a l s o b e e n d o n e i n the cryopreservation field towards the development of xeno-free effective cryopreservation protocols.…”

Section: Cryopreservation In Xeno-free Conditionsmentioning

confidence: 99%

Cryopreservation of Human Pluripotent Stem Cells: Are We Going in the Right Direction?

Martín‐Ibáñez¹,

Hovatta²,

Canals³

2012

Current Frontiers in Cryobiology

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Section: Cryopreservation In Xeno-free Conditionsmentioning

confidence: 99%

Cryopreservation of Human Pluripotent Stem Cells: Are We Going in the Right Direction?

Martín‐Ibáñez¹,

Hovatta²,

Canals³

2012

Current Frontiers in Cryobiology

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“…Human essary elimination of remaining pluripotent cells, beESCs were cultured as previously described (37). cause even a few pluripotent cells may be sufficient to Briefly, hESCs were cultured on top of mitotically inacgive rise to tumors (16).…”

Section: Sundberg Et Almentioning

confidence: 99%

Markers of Pluripotency and Differentiation in Human Neural Precursor Cells Derived from Embryonic Stem Cells and CNS Tissue

et al. 2011

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Cell transplantation therapies for central nervous system (CNS) deficits such as spinal cord injury (SCI) have been shown to be effective in several animal models. One cell type that has been transplanted is neural precursor cells (NPCs), for which there are several possible sources. We have studied NPCs derived from human embryonic stem cells (hESCs) and human fetal CNS tissue (hfNPCs), cultured as neurospheres, and the expression of pluripotency and neural genes during neural induction and in vitro differentiation. mRNA for the pluripotency markers Nanog, Oct-4, Gdf3, and DNMT3b were downregulated during neural differentiation of hESCs. mRNA for these markers was found in nonpluripotent hfNPC at higher levels compared to hESC-NPCs. However, Oct-4 protein was found in hESC-NPCs after 8 weeks of culture, but not in hfNPCs. Similarly, SSEA-4 and CD326 were only found in hESC-NPCs. NPCs from both sources differentiated as expected to cells with typical features of neurons and astrocytes. The expressions of neuronal markers in hESC-NPCs were affected by the composition of cell culture medium, while this did not affect hfNPCs. Transplantation of hESC-NPC or hfNPC neurospheres into immunodeficient mouse testis or subcutaneous tissue did not result in tumor formation. In contrast, typical teratomas appeared in all animals after transplantation of hESC-NPCs to injured or noninjured spinal cords of immunodeficient rats. Our data show that transplantation to the subcutaneous tissue or the testes of immunodeficient mice is not a reliable method for evaluation of the tumor risk of remaining pluripotent cells in grafts.

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“…However, animal-derived components may be contaminated with human pathogens, including viruses and prions [24,25]. In our initial screen, we identified five of 21 plant peptones that enhanced the growth of A. tumefaciens and increased the levels of DsRed transient expression when used to replace the animal-derived components in the original YEB recipe.…”

Section: Cultivation Media Without Animal-derived Components Are Necementioning

confidence: 99%

“…As well as increasing the OD 600 and enhancing transient expression, the exclusion of complex animal-derived ingredients is beneficial from a GMP perspective because the risk of product contamination with human and animal pathogens is lower [24,25]. This may reduce the costs associated with specific quality control tests, e.g.…”

Section: Cultivation Media Without Animal-derived Components Are Necementioning

confidence: 99%

Animal component‐free Agrobacterium tumefaciens cultivation media for better GMP‐compliance increases biomass yield and pharmaceutical protein expression in Nicotiana benthamiana

Houdelet

Galinski

Holland

et al. 2017

Biotechnology Journal

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Transient expression systems allow the rapid production of recombinant proteins in plants. Such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. However, large‐scale transient expression requires the production of recombinant Agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. The complex media often used for the cultivation of this species typically include animal‐derived ingredients that can contain human pathogens, thus conflicting with the requirements of good manufacturing practice (GMP). We replaced all the animal‐derived components in yeast extract broth (YEB) cultivation medium with soybean peptone, and then used a design‐of‐experiments approach to optimize the medium composition, increasing the biomass yield while maintaining high levels of transient expression in subsequent infiltration experiments. The resulting plant peptone Agrobacterium medium (PAM) achieved a two‐fold increase in OD600 compared to YEB medium during a 4‐L batch fermentation lasting 18 h. Furthermore, the yields of the monoclonal antibody 2G12 and the fluorescent protein DsRed were maintained when the cells were cultivated in PAM rather than YEB. We have thus demonstrated a simple, efficient and scalable method for medium optimization that reduces process time and costs. The final optimized medium for the cultivation of A. tumefaciens completely lacks animal‐derived components, thus facilitating the GMP‐compliant large‐scale transient expression of recombinant proteins in plants.

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Testing of nine different xeno-free culture media for human embryonic stem cell cultures

Abstract: None of the studied xeno-free media was able to maintain the undifferentiated growth of hESC. The medium containing 20% human serum was found to sustain undifferentiated hESC proliferation to some extent, yet was inferior to the conventional ko-SR-containing medium.

Cited by 126 publications

References 25 publications

Cryopreservation of Human Pluripotent Stem Cells: Are We Going in the Right Direction?

Cryopreservation of Human Pluripotent Stem Cells: Are We Going in the Right Direction?

Markers of Pluripotency and Differentiation in Human Neural Precursor Cells Derived from Embryonic Stem Cells and CNS Tissue

Animal component‐free Agrobacterium tumefaciens cultivation media for better GMP‐compliance increases biomass yield and pharmaceutical protein expression in Nicotiana benthamiana

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