Attachment and growth of human embryonic stem cells on microcarriers

Phillips, Blaine; Horne, Rachel; Lay, Tan Suat; Rust, William L.; Teck, Tan Thong; Crook, Jeremy Micah

doi:10.1016/j.jbiotec.2008.07.1997

Cited by 154 publications

(82 citation statements)

References 28 publications

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“…Industrializing these therapies will require the development of scalable and controlled suspension bioreactors for PSC‐based cell therapy manufacturing. The main culture methods used to translate laboratory adherent culture (Figure 1a‐i) to suspension‐based expansion of PSC are as cell aggregates in stirred suspension (Figure 1a‐ii; Amit et al, 2011; Kehoe, Jing, Lock, & Tzanakakis, 2010; Zweigerdt, Olmer, Singh, Haverich, & Martin, 2011) or using microcarriers as a substrate for cell attachment (Figure 1a‐iii; Chen, Chen, Choo, Reuveny, & Oh, 2011; Oh et al, 2009; Phillips et al, 2008). Aggregate‐based expansion methods provide simplicity and a reduction in the number of processing steps required.…”

Section: Introductionmentioning

confidence: 99%

Chemically controlled aggregation of pluripotent stem cells

Lipsitz

Tonge

Zandstra

2018

Biotech & Bioengineering

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Heterogeneity in pluripotent stem cell (PSC) aggregation leads to variability in mass transfer and signaling gradients between aggregates, which results in heterogeneous differentiation and therefore variability in product quality and yield. We have characterized a chemical‐based method to control aggregate size within a specific, tunable range with low heterogeneity, thereby reducing process variability in PSC expansion. This method enables controlled, scalable, stirred suspension‐based manufacturing of PSC cultures that are critical for the translation of regenerative medicine strategies to clinical products.

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Section: Introductionmentioning

confidence: 99%

Chemically controlled aggregation of pluripotent stem cells

Lipsitz

Tonge

Zandstra

2018

Biotech & Bioengineering

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“…Commercially available microcarriers-such as Cultisphere S, Cytodex 3, Solohill carriers and Hillex II have been widely used for the expansion of ESC, with Cytodex 3 being the most common. The speed used for mouse ESC, varies between 40 and 70 RPM (Fok and Zandstra 2005;Abranches et al 2007;Alfred et al 2011;Fernandes et al 2007;Marinho et al 2010;Storm et al 2010;Tielens et al 2007), while the range for human ESC lies within 24-80 RPM (Storm et al 2010;Chen et al 2011;Fernandes et al 2009;Kehoe et al 2010;Leung et al 2011;Lock and Tzanakakis 2009;Marinho et al 2013;Nie et al 2009;Oh et al 2009;Phillips et al 2008;Serra et al 2010). iPSC are by nature, delicate, making their large scale culture in spinner flasks using microcarriers a much more difficult proposition.…”

Section: Introductionmentioning

confidence: 99%

Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

et al. 2014

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In recent times, the study and use of induced pluripotent stem cells (iPSC) have become important in order to avoid the ethical issues surrounding the use of embryonic stem cells. Therapeutic, industrial and research based use of iPSC requires large quantities of cells generated in vitro. Mammalian cells, including pluripotent stem cells, have been expanded using 3D culture, however current limitations have not been overcome to allow a uniform, optimized platform for dynamic culture of pluripotent stem cells to be achieved. In the current work, we have expanded mouse iPSC in a spinner flask using Cytodex 3 microcarriers. We have looked at the effect of agitation on the microcarrier survival and optimized an agitation speed that supports bead suspension and iPS cell expansion without any bead breakage. Under the optimized conditions, the mouse iPSC were able to maintain their growth, pluripotency and differentiation capability. We demonstrate that microcarrier survival and iPS cell expansion in a spinner flask are reliant on a very narrow range of spin rates, highlighting the need for precise control of such set ups and the need for improved design of more robust systems.

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“…The other microcarriers tested in this study were of crosslinked polystyrene core instead of modified polystyrene core and coated with different proteins or not coated instead of coated with TMA. Previous study has shown that Hillex® II can be used for efficient propagation of human skin fibroblast (Phillips et al 2008). Microcarriers together with agitated suspension culture systems have been used for culture of anchorage dependent cells for production of virus (as vaccines), growth factors and antibodies (Merten 2015).…”

Section: Discussionmentioning

confidence: 99%

Effects of Secretome from Dynamic 3D Cell Culture System onto Growth and Cytoprotection of Nasal Fibroblast

Lokanathan¹

2017

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ABSTRAK Model tiga dimensi (3D) menyerupai ciri-ciri persekitaran tisu asli, justeru morfologi dan isyarat-isyarat sel daripada kultur 3D selalunya lebih menyerupai fisiologi asal berbanding sel kultur dua dimensi (2D ABSTRACTThree dimensional (3D) models mimic the features of native tissue environment. Thus, morphology and signalling of cells from 3D culture are often more physiological than routine two dimensional (2D) cell culture. It is also known that the cell-secreted products have paracrine effect on other cells growth. In this experimental study, we optimised the nasal fibroblast culture on a 3D cell culture system and studied the effects of secretome from the 3D culture (3DCM) onto fibroblast growth and cytoprotection. Nasal fibroblast was isolated from human nasal turbinates. The suitable microcarrier was selected by culturing the fibroblasts in passage 3 on various types PolyGEM™ polystyrene microcarriers. Then, the cells were cultured on selected microcarrier using a 3D culture system and the conditioned medium (CM) was collected. 3DCM were supplemented to fibroblasts to study for attachment, proliferation, and cytoprotective effects against cytotoxicity of Centella asiatica. Bicinchonic Acid Assay (BCA) was performed to quantify protein amount in CMs. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed for preliminary profiling and comparison of 2DCM and 3DCM protein profile. Our study showed that the 3DCM did not significantly enhance cell attachment and proliferation. The secretome of both 2DCM and 3DCM found to have significant cytoprotective effect onto nasal fibroblast against cytotoxicity of C.asiatica extract. 3DCM had higher protein concentration than 2DCM. SDS-PAGE showed three exclusive proteins in 3DCM and four exclusive proteins in 2DCM. Future study should be conducted on utility of nasal fibroblast secretome on cytoprotection against harmful agents in environment and cytotoxicity of natural products.

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Attachment and growth of human embryonic stem cells on microcarriers

Cited by 154 publications

References 28 publications

Chemically controlled aggregation of pluripotent stem cells

Chemically controlled aggregation of pluripotent stem cells

Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

Effects of Secretome from Dynamic 3D Cell Culture System onto Growth and Cytoprotection of Nasal Fibroblast

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