A practical approach for intracellular protein delivery.

Abstract:

:Protein delivery represents a powerful tool for experiments in live cells including studies of protein-protein interactions, protein interference with blocking antibodies, intracellular trafficking and protein or peptide biological functions. Most available reagents dedicated to the protein delivery allow efficient crossing of the plasma membrane. Nevertheless, the major disadvantage for these reagents is a weak release of the delivered protein into the cytoplasm. In this publication we demonstrate efficient protein delivery with a non-peptide based reagent, in human epithelial carcinoma HeLa cells and primary human skin fibroblasts. Using a fluorescent protein in combination with fluorescence microscopy and fluorescence-assisted cell sorting analysis, we show that the delivered protein is indeed released effectively in the cytoplasm, as expected for a dedicated carrier. Furthermore, we present a step-by-step method to optimize conditions for successful intracellular protein delivery.

journal_name

Cytotechnology

journal_title

Cytotechnology

authors

Weill CO,Biri S,Adib A,Erbacher P

doi

10.1007/s10616-007-9102-3

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

41-8

issue

1

eissn

0920-9069

issn

1573-0778

journal_volume

56

pub_type

杂志文章
  • Interaction ofEscherichia coli cytotoxic necrotizing factor type 1 (CNF1) with cultured cells.

    abstract::The cytotoxic necrotizing factor type 1 (CNF1) fromE. coli causes necrosis in rabbit skin and multinucleation in cultured cells. Cells exposed to CNF1 were characterized by changes in actin organization, mainly consisting in the presence of well-developed stress fibers and membrane ruffles. The interaction of CNF1 wit...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00746055

    authors: Falzano L,Fiorentini C,Boquet P,Donelli G

    更新日期:1993-01-01 00:00:00

  • A simple method for in situ freezing of anchorage-dependent cells including rat liver parenchymal cells.

    abstract::We developed a simple method for freezing anchorage-dependent cells, including primary cultured rat liver parenchymal cells, without detaching the cells from the culture dish. The method consists of preculture of the cells to confluence, changing the growth medium to a conventional freezing medium, packaging in a cont...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00556297

    authors: Ohno T,Saijo-Kurita K,Miyamoto-Eimori N,Kurose T,Aoki Y,Yosimura S

    更新日期:1991-03-01 00:00:00

  • Optimization of cultivation conditions for monoclonal IgM antibody production by M1A2 hybridoma using artificial neural network.

    abstract::Monoclonal antibody (McAb) has been established as one of the most successful therapeutic strategies for the treatment of cancer. M1A2 (McAb) as a new monoclonal antibody was designed to recognize heat shock protein (HSP60), but its optimum production condition has not been studied. In this study, the cell culture con...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-019-00330-5

    authors: Bashokouh F,Abbasiliasi S,Tan JS

    更新日期:2019-07-16 00:00:00

  • Cetyltriethylammonium bromide stimulating transcription of Bombyx mori nucleopolyhedrovirus gp64 gene promoter mediated by viral factors.

    abstract::To characterize the effects of cetyltriethylammonium bromide (CTAB) on the transcription of gp64 promoter from Bombyx mori nucleopolyhedrovirus (BmNPV), the plasmid pBmgp64Luc used in transient expression assay system was constructed by using the luciferase gene as a reporter under the control of BmNPV gp64 promoter. ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1024231023015

    authors: Zhou Y,Yi Y,Zhang Z,He J,Zhang Y

    更新日期:2003-01-01 00:00:00

  • Detection of mycoplasma contaminations by the polymerase chain reaction.

    abstract::The polymerase chain reaction (PCR) has been used for the general detection of Mollicutes. 25 Mycoplasma and Acholeplasma species were detected including important contaminants of cell cultures such as M. orale, M. arginini, M. hyorhinis, M. fermentans, A. laidlawii and additional human and animal mycoplasmas. PCR rea...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00754609

    authors: Wirth M,Berthold E,Grashoff M,Pfützner H,Schubert U,Hauser H

    更新日期:1994-01-01 00:00:00

  • Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus.

    abstract::Understanding the growth dynamics of influenza viruses is an essential step in virus replication and cell-adaptation. The aim of this study was to elucidate the growth kinetic of a low pathogenic avian influenza H9N2 subtype in chicken embryo fibroblast (CEF) and chicken tracheal epithelial (CTE) cells during consecut...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-012-9495-5

    authors: Shahsavandi S,Ebrahimi MM,Mohammadi A,Zarrin Lebas N

    更新日期:2013-05-01 00:00:00

  • High-level expression of a foreign gene by a recombinant baculovirus with an expanded host range.

    abstract::The usefulness of host range expanded viruses as an expressionvector system was investigated by following the expression ofthe E. coli lacZ gene. The host range expanded recombinantviruses were obtained from Sf-21 or BmN-4 cells coinfected withAutographa californica and Bombyx mori nuclearpolyhedrosis viruses. Among t...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008166310368

    authors: Kim HS,Woo SD,Kim WJ,Choi JY,Kang SK

    更新日期:2000-02-01 00:00:00

  • Erythropoietin processing in erythropoietic system and central nervous system.

    abstract::We describe possible functions of carbohydrates attached to growth factors and strategies to examine the functions, concentrating on erythropoietin, a major regulator of erythropoiesis. Erythropoietin in erythropoiesis functions as an endocrine hormone; it is produced by kidney cells and transferred into the circulati...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00744323

    authors: Nagao M,Masuda S,Ueda M,Sasaki R

    更新日期:1995-01-01 00:00:00

  • Cell cycle, cell size and mitochondrial activity of hybridoma cells during batch cultivation.

    abstract::Cell cycle, cell size and rhodamine 123 fluorescence in cell populations of two batch cultures were analysed and quantified with a fluorescence-activated cell sorter (FACS). Two cultures derived from either exponential or stationary phase innocula were investigated in order to demonstrate the dependency of the subsequ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00365929

    authors: al-Rubeai M,Chalder S,Bird R,Emery AN

    更新日期:1991-11-01 00:00:00

  • Improved reproducibility in preparing precision-cut liver tissue slices.

    abstract::Precision-cut liver tissue slices (PCLS) have been used for decades to study pharmacological metabolism as well as toxicology and efficacy of novel substances on primary material under standardized conditions. Slicing of primary liver tissue has been done using different slicing machines. Since there has been great va...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-009-9246-4

    authors: Zimmermann M,Lampe J,Lange S,Smirnow I,Königsrainer A,Hann-von-Weyhern C,Fend F,Gregor M,Bitzer M,Lauer UM

    更新日期:2009-12-01 00:00:00

  • Production of high titre disabled infectious single cycle (DISC) HSV from a microcarrier culture.

    abstract::Disabled Infectious Single Cycle (DISC) HSV-2 has been cultured in the complimentary cell line CR2 to provide high titre bulk material suitable for the purification of the virus as a live viral vaccine. CR2 cells are cultured on the microcarrier Cytodex-1 at 5 g l-1 in small scale (1 l) and larger scale (15 l) reactor...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008005200711

    authors: Zecchini TA,Smith RJ

    更新日期:1999-07-01 00:00:00

  • Knockdown of ribosomal protein S15A inhibits proliferation of breast cancer cells through induction of apoptosis in vitro.

    abstract::To explore the role of ribosomal protein S15A (RPS15A) in breast cancer. The Oncomine database was used to compare the expression of RPS15A in human breast cancer tissues and normal tissues. RPS15A in breast cancer cell line ZR-75-30 and BT474 was specifically knocked down using lentivirus-mediated short hairpin RNAs ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-018-0221-9

    authors: Feng W,Liang C,Wang C,Yu X,Li Q,Yang H

    更新日期:2018-10-01 00:00:00

  • Regulation and expression of transforming growth factor type-beta during early mammalian development.

    abstract::We have examined the effect of differentiation on the expression of different members of the transforming growth factor type-beta (TGF-beta) family using embryonal carcinoma (EC) cells and early mammalian embryos. We determined that TGF-beta activity increases approximately 25-100% when the mouse EC cell line, F9, is ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00563783

    authors: Kelly D,Campbell WJ,Tiesman J,Rizzino A

    更新日期:1990-11-01 00:00:00

  • Insights into protective effects of medium additives on animal cells under fluid stresses: the hydrophobic interactions.

    abstract::Animal cells in suspension culture can suffer severe mechanical damage from bursting gas bubbles or other hydrodynamic force sources. Certain chemical additives in the culture media, particularly some surface-active chemicals, can effectively protect animal cells against such damage. Previously we proposed that the pr...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00353929

    authors: Wu J

    更新日期:1996-01-01 00:00:00

  • Explants of porcine coronary artery in culture: A paradigm for studying the influence of heparin on vascular wall cell proliferation.

    abstract::Explant cultures of porcine coronary artery provided a coculture model, used as a paradigm of arterial wall in contact with vascular prosthesis which allowed the study of spatial and temporal changes in cell phenotype. First cells emerging from the explant had an endothelial phenotype monitored by cytoimmunostaining. ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1016195029359

    authors: Dufresne M,Warocquier-Clérout R

    更新日期:2001-09-01 00:00:00

  • Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells.

    abstract::The Atlantic salmon (Salmo salar) serum lectin (SSL) is a soluble C-type lectin that binds bacteria, including salmon pathogens. This lectin is a cysteine-rich oligomeric protein. Consequently, a Drosophila melanogaster expression system was evaluated for use in expressing SSL. A cDNA encoding SSL was cloned into a ve...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-012-9505-7

    authors: Uribe E,Venkatesan M,Rose DR,Ewart KV

    更新日期:2013-08-01 00:00:00

  • Repeated stimulation by LPS promotes the senescence of DPSCs via TLR4/MyD88-NF-κB-p53/p21 signaling.

    abstract::Dental pulp stem cells (DPSCs), one type of mesenchymal stem cells, are considered to be a type of tool cells for regenerative medicine and tissue engineering. Our previous studies found that the stimulation with lipopolysaccharide (LPS) might introduce senescence of DPSCs, and this senescence would have a positive co...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-017-0180-6

    authors: Feng G,Zheng K,Cao T,Zhang J,Lian M,Huang D,Wei C,Gu Z,Feng X

    更新日期:2018-06-01 00:00:00

  • A modified preplate technique for efficient isolation and proliferation of mice muscle-derived stem cells.

    abstract::We modified an existing protocol to develop a more efficient method to acquire and culture muscle-derived stem cells (MDSCs) and compared the characteristics of cells obtained from the two methods. This method is based on currently used multistep enzymatic digestion and preplate technique. During the replating process...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-018-0262-0

    authors: Xu Z,Yu L,Lu H,Feng W,Chen L,Zhou J,Yang X,Qi Z

    更新日期:2018-12-01 00:00:00

  • Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays.

    abstract::This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC50 and 2×EC50 values were 8 and 16 mg/ml concentrations...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-014-9835-8

    authors: Ciğerci İH,Cenkci S,Kargıoğlu M,Konuk M

    更新日期:2016-08-01 00:00:00

  • Productivity enhancement of recombinant protein in CHO cells via specific promoter activation by oncogenes.

    abstract::To construct a recombinant protein highly producing cell lines, we have previously developed the Oncogene Activated Production (OAP) system by using BHK-21 cells. Here we verified the availability of the OAP system in CHO cells. We firstly generated 'primed' ras amplified CHO cells, ras clone I, by introducing human c...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008048928053

    authors: Katakura Y,Seto P,Miura T,Ohashi H,Teruya K,Shirahata S

    更新日期:1999-09-01 00:00:00

  • Investigation of relationship between IL-6 gene variants and hypertension in Turkish population.

    abstract::Hypertension (HT) is a common and life threating health problem worldwide leading to stroke, heart attack and renal failure. It is characterized by elevated blood pressure forced heart load. Human interleukin-6 (IL-6) and C- reactive protein (CRP) are known to be involved in inflammatory processes. IL-6 gene is a poly...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-014-9732-1

    authors: Karaman E,Urhan Kucuk M,Bayramoglu A,Uzun Göçmen S,Ercan S,Guler HI,Kucukkaya Y,Erden S

    更新日期:2015-12-01 00:00:00

  • A two-compartment cell entrapment bioreactor with three different holding times for cells, high and low molecular weight compounds.

    abstract::A new bioreactor for animal cell cultivation employs two compartments for cells and medium respectively. The two chambers are separated by an ultrafiltration membrane. Cells and solution of collagen or collagen/chitosan mixture were loaded to the cell chamber and were allowed to form gel inside. Contraction of the cel...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00365093

    authors: Scholz M,Hu WS

    更新日期:1990-09-01 00:00:00

  • The role of recombinant proteins in the development of serum-free media.

    abstract::Early developments in serum-free media led to a variety of formulations in which components normally provided in serum and required for growth (insulin, transferrin, lipid supplements, trace elements) and poorly defined components (extracts, hydrolysates) were added to defined basal media. These additives were mostly ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-006-9002-y

    authors: Keenan J,Pearson D,Clynes M

    更新日期:2006-03-01 00:00:00

  • Formation of multilayer rosettes in phytohemagglutinin-stimulated lymphocytes: correlation with chromatin condensation conformation in premature chromosome condensation.

    abstract::We found that the formation of multilayer rosettes by transformed human blood lymphocytes after phytohemagglutinin (PHA) stimulation is correlated with conformational changes of the chromatin as seen by premature chromosome condensation (PCC). The frequency distribution of grades of PCC and multilayer rosette formatio...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00145027

    authors: Thomou H,Sambani C,Kitsiou P,Politis G

    更新日期:1988-07-01 00:00:00

  • Is there any genetic predisposition of MMP-9 gene C1562T and MTHFR gene C677T polymorphisms with essential hypertension?

    abstract::The current study was conducted to determine whether there is a relation between hypertension and two different polymorphisms, including C1562T of the Matrix metalloproteinase-9 (MMP-9) gene and C677T of the methylenetetrahydrofolate reductase (MTHFR) gene. Genomic DNA obtained from 224 persons (125 patients with hype...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-013-9665-0

    authors: Bayramoglu A,Urhan Kucuk M,Guler HI,Abaci O,Kucukkaya Y,Colak E

    更新日期:2015-01-01 00:00:00

  • Comparison of specific rates of hybridoma growth and metabolism in batch and continuous cultures.

    abstract::For the mouse hybridoma cell line VO 208, kinetics of growth, consumption of glucose and glutamine, and production of lactate, ammonia and antibodies were compared in batch and continuous cultures. At a given specific growth rate, different metabolic activities were observed: a 40% lower glucose and glutamine consumpt...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00570891

    authors: Goergen JL,Marc A,Engasser JM

    更新日期:1992-01-01 00:00:00

  • Relative survivability of human osteoblasts is enhanced by 39 °C and ascorbic acid after exposure to photopolymerization ingredients.

    abstract::Photopolymerizable hydrogels offer great potential in cartilage tissue engineering due to their ability to conform to irregular defect shapes and be applied in a potentially minimally invasive manner. An important process requirement in the use of photopolymerizable hydrogels is the ability of the suspended cells to w...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-012-9512-8

    authors: Dua R,Ramaswamy S

    更新日期:2013-08-01 00:00:00

  • Binucleated HeLa cells are formed by cytokinesis failure in starvation and keep the potential of proliferation.

    abstract::Many cytological studies have reported that the numbers of binucleated cells were elevated in various tumors. However, binucleated cells are observed in not only malignant tumors but also normal tissues. Thus, the clinical significance of binucleated cells is controversial. Here we attempted to elucidate the character...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-015-9869-6

    authors: Nishimura K,Watanabe S,Hayashida R,Sugishima S,Iwasaka T,Kaku T

    更新日期:2016-08-01 00:00:00

  • Efficient production of recombinant IgG by metabolic control and co-expression with GLUT5 in a fructose-based medium.

    abstract::A fructose-based cell culture is suitable for the process control of protein production because of slow sugar consumption rate and low lactate accumulation. The fructose transporter, GLUT5, mediates its incorporation into cells and is required for the fructose-based culture. In order to produce efficiently recombinant...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-010-9289-6

    authors: Inoue Y,Tsukamoto Y,Yamanaka M,Nakamura S,Inoue A,Nishino N,Kawahara H

    更新日期:2010-08-01 00:00:00

  • Immunocytochemistry assay in BHK-21 cell line infected with Porcine Sapelovirus.

    abstract::The present study describes an immunocytochemistry (ICC) assay with self-raised hyperimmune sera and a Baby Hamster Kidney-21 (BHK-21) cell line infected with Porcine Sapelovirus (PSV). Sapelovivus/IVRI/SPF-c-6/2015 strain Indian PSV was isolated from the porcine IBRS-2 cell line and investigated for growth on non-por...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-019-00315-4

    authors: Kumari S,Singh R,Desingu PA,Ray PK,Taru Sharma G,Saikumar G

    更新日期:2019-06-01 00:00:00