Combination of baculovirus-mediated gene transfer and rotating-shaft bioreactor for cartilage tissue engineering

Chen, Huang‐Chi; Lee, Hsiao-Ping; Ho, Yi-Chen; Sung, Ming-Lun; Hu, Yu‐Chen

doi:10.1016/j.biomaterials.2006.01.018

Cited by 29 publications

(19 citation statements)

References 36 publications

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“…but disappeared at 24 d.p.t., agreeing with the notion that baculoviral DNA is degraded and diluted within the mammalian cells. 28 To verify the state of the DNA, fluorescent in situ hybridization (FISH) experiment using the egfp probe was performed at 24 d.p.t. Figure 2d illustrates the presence of a significant number of bright fluorescent spots associated with the chromosomes.…”

Section: Comparison Of Orip/ebna1 Aav Itr and Sb Systemsmentioning

confidence: 99%

“…28 The CMV-EGFP cassette was also PCR amplified from pEGFP-N1 (Clontech, Mountain View, CA, USA) and subcloned into the oriP/EBNA1-containing pRep4 (Invitrogen, Carlsbad, CA, USA). The cassette comprising CMV-EGFP and oriP/EBNA1 was then subcloned into pFastBacDpolhDp10, whose polyhedrin and p10 promoters were removed from pFastBac DUAL 20 to yield pBac-COE-CE.…”

Section: Preparation Of Recombinant Baculovirusesmentioning

confidence: 99%

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Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

et al. 2011

Gene Ther

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Antiangiogenesis is an appealing anticancer approach but requires continued presence of the antiangiogenic agents, which can be remedied by gene therapy. Baculovirus is an emerging gene delivery vector but only mediates transient expression (o7 days); thus, this study primarily aimed to develop a hybrid baculovirus for sustained antiangiogenic gene expression and cancer therapy. We first constructed plasmids featuring adeno-associated virus inverted terminal repeats (AAV ITRs), oriP/ Epstein-Barr virus-expressed nuclear antigen 1 (EBNA1) or Sleeping Beauty (SB) transposon and compared their efficacies in terms of persistent expression. In human embryonic kidney (HEK293) cells, AAV ITR failed to prolong the expression while oriP/EBNA1 moderately extended the expression to 35 days. In contrast, the SB system led to stable expression beyond 77 days even without antibiotic selection. Given this finding, we constructed a hybrid SB baculovirus expressing the SB transposase and harboring the transgene cassette flanked by inverted repeat/direct-repeat (IR/DR) elements recognizable by SB. The hybrid SB baculovirus efficiently transduced mammalian cells and mediated an expression duration longer than that by conventional baculoviruses, thanks to the transgene persistence and integration. The SB baculovirus (Bac-SB-T2hEA/w) expressing the antiangiogenic fusion protein comprising endostatin and angiostatin (hEA) also enabled prolonged hEA expression. With sustained hEA expression, Bac-SB-T2hEA/w repressed the angiogenesis in vivo, hindered the growth of two different tumors (prostate tumor allografts and human ovarian tumor xenografts) in mice and extended the life span of animals. These data altogether implicated the potential of the hybrid SB-baculovirus vector for prolonged hEA expression and for the treatment of multiple types of angiogenesis-dependent tumors.

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Section: Comparison Of Orip/ebna1 Aav Itr and Sb Systemsmentioning

confidence: 99%

Section: Preparation Of Recombinant Baculovirusesmentioning

confidence: 99%

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

et al. 2011

Gene Ther

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“…В зависимости от медицинских показаний воз-можно использование хондроцитов, мультипотен-тных мезенхимальных стволовых клеток (ММСК) из различных источников (в основном из костного мозга (КМ) и жировой ткани (ЖТ), клеток-пред-шественников из надкостницы и надхрящницы или генетически-модифицированных клеток [22][23][24][25].…”

Section: технологии стимулирования регенерации поврежденной хрящевunclassified

Tissue Engineering and Regenerative Medicine Technologies in the Treatment of Articular Cartilage Defects

Басок

Севастьянов

2017

RJTAO

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1 ФГБУ «Федеральный научный центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава России, Москва, Российская Федерация 2 АНО «Институт медико-биологических исследований и технологий», Москва, Российская Федерация Важнейшими проблемами здравоохранения в индустриальном обществе являются повреждение и деге-нерация суставного хряща, что связано с ограниченными возможностями ткани к регенерации. В обзоре подробно описаны существующие и разрабатываемые технологии восстановления и замещения повреж-денных хрящевых тканей суставов. Дан анализ полученных результатов по двум основным направлени-ям: стимулирование регенерации поврежденной хрящевой ткани и выращивание элементов хрящевой ткани в биореакторах.Ключевые слова: суставной хрящ, клеточно-и тканеинженерные конструкции, регенеративная медицина, биореакторы. TISSUE ENGINEERING AND REGENERATIVE MEDICINE TECHNOLOGIES IN THE TREATMENT OF ARTICULAR CARTILAGE DEFECTS

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“…18 The mock-transduced and transduced constructs were positioned on the individual needles (eight constructs per reactor). 19 The RSB was oriented horizontally and then half of the reactor space was filled with E100 ml Dulbecco's modified Eagle's medium containing 10% fetal bovine serum, 50 mg ml À1 ascorbic acid and 1% penicillin/ streptomycin/amphotericin B. The reactor was operated in a perfusion mode (medium flow rate ¼ 0.2 ml min À1 , gas flow rate ¼ 20 ml min À1 ) with the shaft rotation speed maintained at 10 r.p.m.…”

Section: Three-dimensional Culturementioning

confidence: 99%

“…18 Furthermore, transduction of chondrocytes with a baculovirus expressing the reporter protein (enhanced green fluorescent protein) obstructs neither chondrocyte differentiation nor the formation of cartilaginous tissues when the chondrocyte/scaffold constructs are cultured in the RSB. 19 In this study, we hypothesized that combining threedimensional (3D) culture in the RSB and baculovirusmediated growth factor expression could ameliorate the growth of cartilaginous tissues. Therefore, we genetically modified the rabbit articular chondrocytes with Bac-CB, a recombinant baculovirus expressing bone morphogenetic protein-2 (BMP-2), seeded the cells into poly (L-lactide-co-glycolide) (PLGA) scaffolds and cultured the constructs in the RSB for 3 weeks.…”

Section: Introductionmentioning

confidence: 99%

Combination of baculovirus-expressed BMP-2 and rotating-shaft bioreactor culture synergistically enhances cartilage formation

et al. 2007

Gene Ther

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Baculovirus is an emerging gene delivery vector, thanks to a number of unique advantages. Herein, we genetically modified the rabbit articular chondrocytes with a recombinant baculovirus (Bac-CB) encoding bone morphogenetic protein-2 (BMP-2), which conferred high level BMP-2 expression and triggered the re-differentiation of dedifferentiated third passage (P3) chondrocytes in the monolayer culture. The transduced and mock-transduced P3 cells were seeded into porous scaffolds and cultured in either the dishes or the rotating-shaft bioreactor (RSB), a novel bioreactor imparting a dynamic, two-phase culture environment. Neither mocktransduced constructs in the RSB culture nor the Bac-CBtransduced constructs in the static culture grew into uniform cartilaginous tissues. Only the Bac-CB-transduced constructs cultured in the RSB for 3 weeks resulted in cartilaginous tissues with hyaline appearance, uniform cell distribution, cartilage-specific gene expression and considerably enhanced cartilage-specific extracellular matrix deposition, as determined by histological staining, reverse transcription-PCR analyses and biochemical assays. This is the first study demonstrating that combination of baculovirusmediated growth factor expression and RSB culture synergistically enhanced in vitro creation of cartilaginous tissues from dedifferentiated chondrocytes. Since baculovirus transduction is generally considered safe, this approach represents a viable alternative to stimulate the formation of engineered cartilage in a more cost-effective way than the growth factor supplementation.

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Combination of baculovirus-mediated gene transfer and rotating-shaft bioreactor for cartilage tissue engineering

Cited by 29 publications

References 36 publications

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

Development of the hybrid Sleeping Beauty-baculovirus vector for sustained gene expression and cancer therapy

Tissue Engineering and Regenerative Medicine Technologies in the Treatment of Articular Cartilage Defects

Combination of baculovirus-expressed BMP-2 and rotating-shaft bioreactor culture synergistically enhances cartilage formation

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