Cell-based high-throughput proliferation and cytotoxicity assays for screening traditional Chinese herbal medicines

Ding, Li; Zang, Ru; Yang, Shang‐Tian; Wang, Jufang; Wang, Xiaoning

doi:10.1016/j.procbio.2013.02.005

Cited by 17 publications

(12 citation statements)

References 30 publications

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“…Microbioreactor array with 3-D fibrous matrix Mouse ESC High-through cell-based assay for drug screening [14,87,102,106] Microbioreactor array with full factorial design of growth factor combinations Human ESC Screening exogenous and paracrine factors in human ESC differentiation into mesoderm cells…”

Section: Understanding Of Cell Migration and Cancer Invasion [104]mentioning

confidence: 99%

“…This 3-D model more resembled the in vivo microenvironment and thus the cell response to the drugs more reflected the response that could occur in vivo. For example, various Chinese herb medicines were tested in the 3-D microbioreactors seeded with mouse ESCs and cancer cells [106] , in [100] .…”

Section: Applications In Stem Cell Differentiation and Drug Screeningmentioning

confidence: 99%

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Engineering stem cell niches in bioreactors

Liu

Liu²,

Zang³

et al. 2013

WJSC

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Stem cells, including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells and amniotic fluid stem cells have the potential to be expanded and differentiated into various cell types in the body. Efficient differentiation of stem cells with the desired tissue-specific function is critical for stem cell-based cell therapy, tissue engineering, drug discovery and disease modeling. Bioreactors provide a great platform to regulate the stem cell microenvironment, known as "niches", to impact stem cell fate decision. The niche factors include the regulatory factors such as oxygen, extracellular matrix (synthetic and decellularized), paracrine/ autocrine signaling and physical forces (i.e. , mechanical force, electrical force and flow shear). The use of novel bioreactors with precise control and recapitulation of niche factors through modulating reactor operation parameters can enable efficient stem cell expansion and differentiation. Recently, the development of microfluidic devices and microbioreactors also provides powerful tools to manipulate the stem cell microenvironment by adjusting flow rate and cytokine gradients. In general, bioreactor engineering can be used to better modulate stem cell niches critical for stem cell expansion, differentiation and applications as novel cell-based biomedicines. This paper reviews important factors that can be more precisely controlled in bioreactors and their effects on stem cell engineering.

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Section: Understanding Of Cell Migration and Cancer Invasion [104]mentioning

confidence: 99%

Section: Applications In Stem Cell Differentiation and Drug Screeningmentioning

confidence: 99%

Engineering stem cell niches in bioreactors

Liu

Liu²,

Zang³

et al. 2013

WJSC

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“…To reduce possible disturbance of Gls, a lower concentration of 0.01 % (w/v) is considered as the optimal dosage in this work. Specific growth rate (l) is an accurate and sensitive parameter for evaluating animal cell proliferation and has been used to assess the effects of traditional Chinese herbal medicines on mouse embryonic stem (mES) cells [34,35]. This work also used this parameter to determine the effect of Gls extract on CHO cells.…”

Section: Growth Stimulating Effect Of Gls Extract On Cho K1 Cellmentioning

confidence: 99%

“…Although the extract of Gls increased the expansion fold of cells cultured in 0 % FBS growth medium, the promotion effect is not strong enough to stop cell number reduction. In previous study with mES cells, high dosage of Gls extract showed significant inhibition on cell growth [34], because of the accumulation of harmful components from the rough-wrought Gls extract [39]. Therefore, it is necessary to determine the optimal dose when applying Gls extract for CHO K1 cell cultures.…”

Section: Growth Stimulating Effect Of Gls Extract On Cho K1 Cellmentioning

confidence: 99%

“…For example, water extracts from Gls can enhance immune functions by activating the proliferation of lymphocytes [31,32] and accelerating the recovery of natural killer (NK) cells [33]. Our previous studies have found that the water-soluble extracts of Gls were able to promote the specific growth rate of mouse embryonic stem (mES) cells, which might possibly be protected by those Gls bioactive compounds against cell apoptosis, and we have successfully used Gls extracts as a medium supplement for scale-up stem cell cultures [34,35].…”

Section: Introductionmentioning

confidence: 99%

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Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation

Ding

Zhong

et al. 2016

Bioprocess Biosyst Eng

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In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production.

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An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

Zang

Xin

Zhang

et al. 2019

Biotech & Bioengineering

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Embryonic stem cell test (EST) is the only generally accepted in vitro method for assessing embryotoxicity without animal sacrifice. However, the implementation and application of EST for regulatory embryotoxicity screening are impeded by its technical complexity, long testing period, and limited endpoint data. In this study, a high throughput embryotoxicity screening based on mouse embryonic stem cells (mESCs) expressing enhanced green fluorescent protein (EGFP) driven by a human survivin promoter and a human cytomegalovirus promoter, respectively, was developed. These EGFP expressing mESCs were cultured in three-dimensional (3D) fibrous scaffolds in microbioreactors on a multiwell plate with EGFP fluorescence signals as cell responses to chemicals monitored noninvasively in a high throughput manner. Nine chemicals with known developmental toxicity were used to validate the survivin-based embryotoxicity assay, which showed that strongly embryotoxic compounds such as 5-fluorouracil, retinoic acid, and methotrexate downregulated survivin expression by more than 50% in 3 days, while weakly embryotoxic compounds such as boric acid, methoxyacetic acid, and tetracyclin showed modest downregulation effect and nonembryotoxic saccharin, penicillin G, and acrylamide had negligible downregulation effect on survivin expression, confirming that survivin can be used as a molecular endpoint for high throughput screening of embryotoxicants. The potential developmental toxicity of three Chinese herbal medicines were also evaluated using this assay, demonstrating its application in in vitro developmental toxicity test for drug safety assessment.

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Cell-based high-throughput proliferation and cytotoxicity assays for screening traditional Chinese herbal medicines

Cited by 17 publications

References 30 publications

Engineering stem cell niches in bioreactors

Engineering stem cell niches in bioreactors

Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

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