<i>In Vitro</i>Effect of Gelatins on Murine Cell Proliferation

Kojima, Takashi; Koide, Tatsurou; Nagata, Hiroshi; Paeng, Noriko; Sano, Mitsuru; Sasanabe, Ryujiro; Horikoshi, Ichiro; Ito, Nobuhiro; Hasegawa, Makoto

doi:10.1089/108497801753354339

Cited by 9 publications

(8 citation statements)

References 6 publications

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“…The high proliferation rate of miPS cells on the hydrogels and Gelatin‐PS scaffold indicates that an undifferentiated state of miPS cells can be well maintained on scaffolds since the undifferentiated stem cells exhibited high proliferation potential, which decreases as stem cells mature and differentiate (Smith et al ., ; Williams et al ., ). It is well known that a Gelatin‐PS scaffold provides excellent cell adhesion and proliferation for mES cells (Yang et al ., , ; Kojima et al ., ). Our results indicate that the two hydrogels had even better performance than the Gelatin‐PS scaffold, especially the PNaAMPS hydrogels.…”

Section: Resultsmentioning

confidence: 97%

Hydrogels as feeder-free scaffolds for long-term self-renewal of mouse induced pluripotent stem cells

Yang

Liu

Kurokawa

et al. 2012

J Tissue Eng Regen Med

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Expanding undifferentiated induced pluripotent stem (iPS) cells in vitro is a basic requirement for application of iPS cells in both fundamental research and clinical regeneration. In this study, we intended to establish a simple, low cost and efficient method for the long-term self-renewal of mouse induced pluripotent stem (miPS) cells without using feeder-cells and adhesive proteins. Three scaffolds were selected for the long-term subculture of miPS cells over two months starting from passages 14 to 29: 1) a gelatin coated polystyrene (Gelatin-PS) that is a widely used scaffold for self-renewal of mouse embryonic stem (mES) cells; 2) a neutral hydrogel poly(N,N-dimethylacrylamide) (PDMAAm); and 3) a negatively charged hydrogel poly(2-acrylamido-2-methyl-propane sulfonic acid sodium salt) (PNaAMPS). Each passaged miPS cells on these scaffolds were cryopreserved successfully and the revived cells showed high viability and proliferation. The passaged miPS cells maintained a high undifferentiated state on all three scaffolds and a high level of pluripotency by expressing differentiation markers in vitro and forming teratomas in SCID mice with derivatives of all three germ layers. Compared to Gelatin-PS, the two hydrogels exhibited much better self-renewal performance in terms of high proliferation rate and level of expression of undifferentiated gene markers as well as efficiency in pluripotent teratoma formation. Furthermore, the PNaAMPS hydrogel demonstrated a slightly higher efficiency and simpler operation of cell expansion than the PDMAAm hydrogel. To conclude, PNaAMPS hydrogel is an excellent feeder-free scaffold because of its simplicity, low cost and high efficiency in expanding a large number of miPS cells in vitro.

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Section: Resultsmentioning

confidence: 97%

Hydrogels as feeder-free scaffolds for long-term self-renewal of mouse induced pluripotent stem cells

Yang

Liu

Kurokawa

et al. 2012

J Tissue Eng Regen Med

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“…3 MMC-treated spleen cells as well as MMC-treated RL? 1 cells partly suppressed the anti-proliferative activity of PS gelatin to RL?…”

Section: Discussionmentioning

confidence: 99%

“…2 Afterward, we observed that BB gelatin stimulates proliferation of murine spleen cells, especially T cells 2 and that porcine skin (PS) gelatin inhibits proliferation of several kinds of murine tumor cells followed by apoptosis of those tumor cells. 3 In this study, we used two kinds of gelatin (BB and PS gelatins) and two sorts of murine cells (spleen cells of Balb/c mice and the malignant counterpart, RL? 1 cells) in order to investigate two contrary actions of PS and BB gelatins on murine benign and malignant cells.…”

Section: Introductionmentioning

confidence: 99%

Comparison between Bovine Bone and Porcine Skin Gelatins in their Effects on Murine Cell Proliferation

Koide

Nagata²,

Kojima³

et al. 2002

Cancer Biotherapy and Radiopharmaceuticals

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We investigated relationship between porcine skin (PS) and bovine bone (BB) gelatins in their actions on proliferation of murine benign and malignant cells in this study. We previously observed that BB gelatin enhanced spleen cell proliferation. The present study showed that such an activity of BB gelatin was not exerted in a serum-free medium. On the other hand, PS gelatin suppressed proliferation of normal spleen cells and of those stimulated by concanavalin A (Con A). It is not known whether or not such an activity of PS gelatin on Con A-stimulated spleen cells is exerted in the serum-free medium since Con A was unable to augment proliferation of spleen cells in that medium. BB gelatin as well as PS gelatin suppressed proliferation of RL male symbol 1 cells, a T cell lymphoma cell line of Balb/c mice, and such an activity of BB gelatin was not exerted in the serum-free medium whereas PS gelatin exerted its activity in the same medium. Mitomycin C (MMC)-treated spleen cells as well as MMC-treated RL male symbol 1 cells partly released RL male symbol 1 cells from the inhibition of proliferation by PS gelatin. MMC-treated RL male symbol 1 cells as well as MMC-treated spleen cells suppressed the proliferation of spleen cells augmented by BB gelatin. Inhibition of RL male symbol 1 cell proliferation by PS gelatin was not affected by BB gelatin, but enhancement of spleen cell proliferation by BB gelatin was attenuated by PS gelatin regardless of the sequence of treating the spleen cells with PS gelatin. Enhancement of spleen cell proliferation by BB gelatin was time-dependent but suppression of RL male symbol 1 cell proliferation by PS gelatin was not. In conclusion, BB gelatin enhanced proliferation of spleen cells and suppressed proliferation of RL male symbol 1 cells. In both cases, fetal calf serum (FCS) was required. PS gelatin suppressed proliferation of spleen cells and of RL male symbol 1 cells without FCS.

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“…For this study, we used equine bone marrow-derived (BM-)MSCs, because the horse is both a patient with a large clinical need for better articular cartilage repair and a recommended animal model for pre-clinical testing of cartilage regeneration strategies 50,51 . Further, equine MSCs do express the receptor that allows endotoxin signalling, namely TLR4 52,53 , and have previously been shown to be sensitive to LPS 21 . Our data suggest that the presence of endogenous LPS does, however, not affect chondrogenic differentiation of healthy equine BM-MSCs embedded within GelMA hydrogels.…”

Section: Effect Of Endotoxinsmentioning

confidence: 99%

“…GelMAs in the production of collagen type I. Gelatine types A and B are produced in different ways 59 , which results in different isoelectric points 60 and can influence degree of methacrylation and molecular weight 59 . Furthermore, the species and tissue type from which the gelatine is extracted could influence cell response 52,53 . Type A and B variants from bovine and porcine origin were included, but as the current study was not designed to identify the influence of gelatine type, further research is needed to elucidate which qualities of type A and type B GelMA could influence MSC differentiation and thereby impact tissue engineering and regenerative medicine applications.…”

Section: Effect Of Gelatine Characteristicsmentioning

confidence: 99%

Impact of endotoxins in gelatine hydrogels on chondrogenic differentiation and inflammatory cytokine secretionin vitro

Groen

Utomo

Castilho

et al. 2020

Preprint

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Gelatine methacryloyl (GelMA) hydrogels are widely used in studies aiming at cartilage regeneration. However, the endotoxin content of commercially available GelMAs and gelatines used in these studies is often overlooked, even though endotoxins may influence several cellular functions. Moreover, regulations for clinical use of biomaterials dictate a stringent endotoxin limit.We determined the endotoxin level of five different GelMAs and evaluated the effect on the chondrogenic differentiation of equine mesenchymal stromal cells (MSCs). Cartilage-like matrix production was evaluated by biochemical assays and immunohistochemistry. Furthermore, equine peripheral blood mononuclear cells (PBMCs) were cultured on the hydrogels for 24 hours, followed by the assessment of tumour necrosis factor (TNF)-α and C-C motif chemokine ligand (CCL)2 as inflammatory markers.The GelMAs were found to have widely varying endotoxin content (two with >1000 EU/ml and three with <10 EU/ml), however, this was not a critical factor determining in vitro cartilage-like matrix production of embedded MSCs. PBMCs did produce significantly higher TNF-α and CCL2 in response to the GelMA with the highest endotoxin level compared to the other GelMAs.Although limited effects on chondrogenic differentiation were found in this study, caution with the use of commercial hydrogels is warranted in the translation from in vitro to in vivo studies because of regulatory constraints and potential inflammatory effects of the content of these hydrogels.

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In VitroEffect of Gelatins on Murine Cell Proliferation

Cited by 9 publications

References 6 publications

Hydrogels as feeder-free scaffolds for long-term self-renewal of mouse induced pluripotent stem cells

Hydrogels as feeder-free scaffolds for long-term self-renewal of mouse induced pluripotent stem cells

Comparison between Bovine Bone and Porcine Skin Gelatins in their Effects on Murine Cell Proliferation

Impact of endotoxins in gelatine hydrogels on chondrogenic differentiation and inflammatory cytokine secretionin vitro

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