Tissue Technology Inc.

Miami, FL, United States

Tissue Technology Inc.

Miami, FL, United States
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Chen H.-C.,Tissue Technology Inc. | Chen H.-C.,Chang Gung Memorial Hospital | Chen H.-C.,Chang Gung University | Zhu Y.-T.,Tissue Technology Inc. | And 2 more authors.
PLoS ONE | Year: 2012

Contact-inhibition ubiquitously exists in non-transformed cells and explains the poor regenerative capacity of in vivo human retinal pigment epithelial cells (RPE) during aging, injury and diseases. RPE injury or degeneration may unlock mitotic block mediated by contact inhibition but may also promote epithelial-mesenchymal transition (EMT) contributing to retinal blindness. Herein, we confirmed that EMT ensued in post-confluent ARPE-19 cells when contact inhibition was disrupted with EGTA followed by addition of EGF and FGF-2 because of activation of canonical Wnt and Smad/ZEB signaling. In contrast, knockdown of p120-catenin (p120) unlocked such mitotic block by activating p120/Kaiso, but not activating canonical Wnt and Smad/ZEB signaling, thus avoiding EMT. Nuclear BrdU labeling was correlated with nuclear release of Kaiso through p120 nuclear translocation, which was associated with activation of RhoA-ROCK signaling, destabilization of microtubules. Prolonged p120 siRNA knockdown followed by withdrawal further expanded RPE into more compact monolayers with a normal phenotype and a higher density. This new strategy based on selective activation of p120/Kaiso but not Wnt/β-catenin signaling obviates the need of using single cells and the risk of EMT, and may be deployed to engineer surgical grafts containing RPE and other tissues. © 2012 Chen et al.


Tseng S.C.G.,Tissue Technology Inc. | Chen S.-Y.,Tissue Technology Inc. | Shen Y.-C.,National Taiwan University Hospital | Chen W.-L.,National Taiwan University Hospital | Hu F.-R.,National Taiwan University Hospital
Current Molecular Medicine | Year: 2010

The stem cells (SCs) of the corneal epithelium located in the limbal basal layer are the ultimate source to maintain corneal epithelial homeostasis. Like other adult tissue-specfic SCs, self renewal and fate decision of limbal SCs are regulated by a specialized in vivo microenvironment, termed "niche". Loss of limbal SCs or dysfunction of the limbal niche renders corneas with a unique clinical disease labeled limbal stem cell deficiency (LSCD). Besides transplantation of autologous or allogeneic limbal SCs or amniotic membrane, a new strategy of treating LSCD is to transplant a bio-engineered graft by expanding limbal SCs ex vivo. Herein, we conduct a critical appraisal of six protocols that have successfully been practiced in treating human patients with LSCD, and identify issues whether niche regulation has been disrupted or maintained during isolation and expansion. Consequently, we propose a future direction that may circumvent the potential pitfalls existing in these conventional protocols by preserving the interaction between limbal SCs and their native niche cells during isolation and expansion. Such an approach may one day help realize considerable promise held by adult SCs in treating a number of diseases. © 2010 Bentham Science Publishers Ltd.


PubMed | Huazhong University of Science and Technology, Dalian Central Hospital, Shanghai JiaoTong University and Tissue Technology Inc.
Type: | Journal: Scientific reports | Year: 2016

Human corneal endothelial cells (HCECs) have limited proliferative capacity due to contact-inhibition at G1 phase. Such contact-inhibition can be delayed from Day 21 to Day 42 by switching EGF-containing SHEM to LIF/bFGF-containing MESCM through transient activation of LIF-JAK1-STAT3 signaling that delays eventual nuclear translocation of p16


Zhu Y.-T.,Tissue Technology Inc | Han B.,Huazhong University of Science and Technology | Li F.,Shandong University | Chen S.-Y.,Tissue Technology Inc | And 3 more authors.
Investigative Ophthalmology and Visual Science | Year: 2014

PURPOSE. To determine the signaling pathway involved in expanding contact-inhibited human corneal endothelial cells (HCECs) using p120 and Kaiso small interfering RNAs (siRNAs). METHODS. Expansion of HCEC monolayers on collagen IV in SHEM using p120 siRNA was optimized regarding various dosage, frequency, and starting date before being added Kaiso siRNA or various inhibitors of Rho, ROCK, NFjB, and TAK1. Phase contrast micrographs were used for monitoring cell shape, monolayer size, and cell density. Immunostaining was used to determine cytolocalization of BrdU, p120, pNFkB, F-actin, a-catenin, b-catenin, LEF1, Na{thorn}/ K{thorn}-ATPase, N-cadherin, ZO-1, and S100A4. Western blotting was used to determine the protein level of RhoA and RhoA-guanosine-50-triphosphate (GTP). RESULTS. The HCEC monolayer size in diameter was expanded from 2.1 6 0.4 mm to 4.3 6 0.3 mm (P < 0.05) by increasing p120 siRNA from 40 nM to 100 nM starting at day 7, to 5.0 6 0.4 mm (P < 0.05) by adding 100 nM Kaiso siRNA, to 6.8 6 0.3 mm by using one-fourth corneoscleral rim (P < 0.05), and to 8.1 6 0.5 mm by using one-half corneoscleral rim (P < 0.05). Such proliferative effect required activation of RhoA-ROCK-noncanonical bone morphogenic protein (BMP) signaling and nuclear translocation of phosphorylated nuclear factor kappa-light-chain-enhancer of activated B cells (pNFjB). After withdrawal of siRNAs for 1 week, the resultant HCEC monolayer maintained a hexagonal shape, the average cell density of 2254 6 87 mm2 (n 1/4 3), and normal expression patterns of F-actin, a-catenin, b-catenin, Ncadherin, ZO-1, and Na{thorn}/K{thorn}-ATPase without S100A4 and alpha-smooth muscle actin (a-SMA). CONCLUSIONS. The optimized knockdown with p120 and Kaiso siRNAs further expands the size of HCEC monolayers without endothelial mesenchymal transition (EMT) via selective activation of p120/Kaiso signaling that requires the RhoA-ROCK-noncanonical BMP-NFkB signaling. © 2014 The Association for Research in Vision and Ophthalmology, Inc.


Zhu Y.,Arizona Cancer Center | Zhu Y.,Tissue Technology Inc. | Zhu M.,Arizona Cancer Center | Lance P.,Arizona Cancer Center
Biochemical and Biophysical Research Communications | Year: 2012

COX-2 is a major regulator implicated in colonic cancer. However, how COX-2 signaling affects colonic carcinogenesis at cellular level is not clear. In this article, we investigated whether activation of COX-2 signaling by deoxycholic acid (DCA) in primary human normal and cancer associated fibroblasts play a significant role in regulation of proliferation and invasiveness of colonic epithelial cancer cells. Our results demonstrated while COX-2 signaling can be activated by DCA in both normal and cancer associated fibroblasts, the level of activation of COX-2 signaling is significantly greater in cancer associated fibroblasts than that in normal fibroblasts. In addition, we discovered that the proliferative and invasive potential of colonic epithelial cancer cells were much greater when the cells were co-cultured with cancer associated fibroblasts pre-treated with DCA than with normal fibroblasts pre-treated with DCA. Moreover, COX-2 siRNA attenuated the proliferative and invasive effect of both normal and cancer associate fibroblasts pre-treated with DCA on the colonic cancer cells. Further studies indicated that the activation of COX-2 signaling by DCA is through protein kinase C signaling. We speculate that activation of COX-2 signaling especially in cancer associated fibroblasts promotes progression of colonic cancer. © 2012 Elsevier Inc.


Zhu Y.,Arizona Cancer Center | Zhu Y.,Tissue Technology Inc | Zhu M.,Arizona Cancer Center | Lance P.,Arizona Cancer Center
Experimental Cell Research | Year: 2012

COX-2 is a major inflammatory mediator implicated in colorectal inflammation and cancer. However, the exact origin and role of COX-2 on colorectal inflammation and carcinogenesis are still not well defined. Recently, we reported that COX-2 and iNOS signalings interact in colonic CCD18Co fibroblasts. In this article, we investigated whether activation of COX-2 signaling by IL1Β in primary colonic fibroblasts obtained from normal and cancer patients play a critical role in regulation of proliferation and invasiveness of human colonic epithelial cancer cells. Our results demonstrated that COX-2 level was significantly higher in cancer associated fibroblasts than that in normal fibroblasts with or without stimulation of IL-1Β, a powerful stimulator of COX-2. Using in vitro assays for estimating proliferative and invasive potential, we discovered that the proliferation and invasiveness of the epithelial cancer cells were much greater when the cells were co-cultured with cancer associated fibroblasts than with normal fibroblasts, with or without stimulation of IL1Β. Further analysis indicated that the major COX-2 product, prostaglandin E2, directly enhanced proliferation and invasiveness of the epithelial cancer cells in the absence of fibroblasts. Moreover, a selective COX-2 inhibitor, NS-398, blocked the proliferative and invasive effect of both normal and cancer associate fibroblasts on the epithelial cancer cells, with or without stimulation of IL-1Β. Those results indicate that activation of COX-2 signaling in the fibroblasts plays a major role in promoting proliferation and invasiveness of the epithelial cancer cells. In this process, PKC is involved in the activation of COX-2 signaling induced by IL-1Β in the fibroblasts. © 2012 Elsevier Inc.


Zhu Y.,Arizona Cancer Center | Zhu Y.,Tissue Technology Inc | Zhu M.,Arizona Cancer Center | Lance P.,Arizona Cancer Center
Experimental Cell Research | Year: 2012

COX-2 and iNOS are two major inflammatory mediators implicated in colorectal inflammation and cancer. Previously, the role of colorectal fibroblasts involved in regulation of COX-2 and iNOS expression was largely ignored. In addition, the combined interaction of COX-2 and iNOS signalings and their significance in the progression of colorectal inflammation and cancer within the fibroblasts have received little investigation. To address those issues, we investigated the role of colonic fibroblasts in the regulation of COX-2 and iNOS gene expression, and explored possible mechanisms of interaction between COX-2 and iNOS signalings using a colonic CCD-18Co fibroblast line and LPS, a potential stimulator of COX-2 and iNOS. Our results clearly demonstrated that LPS activated COX-2 gene expression and enhanced PGE 2 production, stimulated iNOS gene expression and promoted NO production in the fibroblasts. Interestingly, activation of COX-2 signaling by LPS was not involved in activation of iNOS signaling, while activation of iNOS signaling by LPS contributed in part to activation of COX-2 signaling. Further analysis indicated that PKC plays a major role in the activation and interaction of COX-2 and iNOS signalings induced by LPS in the fibroblasts. © 2012 Elsevier Inc.


Li F.,Shandong University | Zhu Y.-T.,Tissue Technology Inc.
Cellular Signalling | Year: 2015

COX-2 is a major regulator in colorectal inflammation and cancer. Herein, we first report that primary cancer-associated colonic fibroblasts activated by HGF play a critical role in mediation of proliferation and invasiveness of human colonic epithelial cancer cells. We have discovered that the proliferation and invasiveness of colonic epithelial cancer cells are predominantly enhanced through activation of PKC-cMET-ERK1/2-COX-2 signaling by HGF in the co-cultured cancer-associated fibroblasts. This conclusion is supported by the fact, that a selective PKC inhibitor, BIM, inhibits ERK1/2 and COX-2 signalings, MEK/ERK1/2 inhibitor, PD98059, nullifies COX-2 signaling, and COX-2 inhibitor, NS-398, attenuates the proliferation and invasiveness potential of the colonic cancer cells. We have concluded that HCF-activated cancer associated fibroblasts play a critical role in carcinogenesis of colonic cancer. © 2015 Elsevier Inc.


Zhu M.,Arizona Cancer Center | Zhu Y.,Arizona Cancer Center | Zhu Y.,Tissue Technology Inc | Lance P.,Arizona Cancer Center
Cell Proliferation | Year: 2013

Objectives: Up to now it has been unclear whether stromal/epithelial interaction affects progression of colon cancer. This study was designed to examine effects of tumour necrosis factor alpha (TNFα)-activated stromal cyclooxygenase-2 (COX-2) signalling on proliferation and invasiveness of colon cancer epithelial cells. Materials and methods: Cyclooxygenase-2 mRNA and protein were determined by real-time PCR and western blotting and prostaglandin E2 (PGE2) was assayed by radioimmunoassay. Cell proliferation and invasiveness were determined by transwell chamber assays and protein kinase C (PKC) was assayed by Biotrak™ PKC Assay System. Results: Our results indicated that TNFα, a powerful inflammatory cytokine, strongly promoted COX-2 expression and PGE2 production in colon cancer-associated fibroblasts. Using in vitro assays for estimating proliferative and invasive potential, we discovered that activation of stromal COX-2 signalling significantly promoted proliferation and invasiveness of colon cancer epithelial cells. In addition, selective COX-2 inhibitor N-[2-(Cyclohexyloxy)-4-nitrophenyl]methanesulfonamide, blocked such proliferative and invasive effects on the cancer epithelial cells. In this process, PKC was involved in activation of COX-2 signalling in the fibroblasts. Conclusion: We conclude that activation of stromal COX-2 signalling by TNFα played a major role in promoting proliferation and invasiveness of colon cancer epithelial cells. © 2013 John Wiley & Sons Ltd.


PubMed | The Second Peoples Hospital of Yunnan Province, Nanjing Medical University, Electronic Technology Inc., YanAn Hospital of Kunming City and Tissue Technology Inc.
Type: | Journal: Clinical colorectal cancer | Year: 2016

Colorectal cancer is a major worldwide health care problem that accounts for 1 million new cases each year. The risk factors for this disease include hereditary factors, environmental agents, and inflammatory stimuli that affect the gastrointestinal tract. Among these risk factors, cyclooxygenase-2 (COX-2) is one of the major players in the progression of colorectal cancer; however, the detailed mechanism of its role in causing colorectal cancer is still not well understood. In addition, the role of COX-2 signaling through the interaction in the epithelial and stromal compartments on colorectal carcinogenesis has not been fully illustrated. In the present review, we provide published evidence to demonstrate that (1) COX-2 signaling plays a major role in the progression of colorectal cancer, (2) activation of COX-2 in the stromal compartment also contributes to colorectal carcinogenesis, and (3) inhibition of COX-2 signaling by COX-2 inhibitors might be an effective method to control colorectal cancer. We have also summarized recent advances and insights from mechanistic studies of colorectal cancer to help prevent and control this deadly disease and provide our opinion regarding the importance of risk reduction and disease prevention for colorectal cancer.

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