Quantitative study of molecular transport due to electroporation: uptake of bovine serum albumin by erythrocyte ghosts

Prausnitz, Mark R.; Milano, C.D.; Gimm, J.A.; Langer, Robert; Weaver, James C.

doi:10.1016/s0006-3495(94)80943-9

Cited by 59 publications

(33 citation statements)

References 48 publications

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“…Predictions of peak transfection conditions for bacteria and yeast 22, [44][45][46][47][48] (MAPE = 48 ± 62%) were not as good as for mammalian cells, but predictions of 50% viability conditions were similarly accurate (MAPE = 21 ± 8%) (eg Figure 2d). Finally, we validated predictions by equation 1 for uptake of small compounds and macromolecules [11][12][13]28 (eg Figure 2f). For these studies that predicted the absolute level of molecular transport using five different molecules and three cell types, the overall correlation MAPE value was 101 ± 118%, which shows some discrepancy when compared with the experimental MAPE of 40%.…”

Section: Mammalian Viability Predictionsmentioning

confidence: 87%

“…To develop mathematical expressions that predict electroporation-mediated uptake, loss of viability and gene transfection, we were guided by experimental observations [10][11][12][13] and theoretical calculations. 14,15 Combined, these suggest that molecular uptake has three important features: (1) uptake occurs only above a quasi-threshold field strength, which depends on pulse length and number; (2) uptake then increases with field strength; and (3) at large field strengths, uptake plateaus at a level which depends on pulse length and number.…”

Section: Development Of Predictive Correlationmentioning

confidence: 99%

“…Based on the hypothesis that electroporation occurs universally in cell membranes under conditions largely independent of the detailed composition of those membranes and further bolstered by widespread experimental demGene Therapy onstration of electroporation in a diversity of cell types, [1][2][3][4] we tested the ability of equations 1 and 2 to predict uptake of a range of different molecules (mostly DNA) and viability for over 900 experimental conditions, including data from 60 different cell lines from 33 different literature studies (mostly in vitro, some in vivo) [11][12][13] ( Figure 2). Good agreement between predictions and this large and diverse set of data would (1) provide a broadly applicable and quantitative means to analyze and optimize electroporation protocols and (2) support the above hypothesis regarding electroporation universality.…”

Section: Validation Of Predictionsmentioning

confidence: 99%

See 2 more Smart Citations

Prediction and optimization of gene transfection and drug delivery by electroporation

Canatella

Prausnitz

2001

Gene Ther

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Section: Mammalian Viability Predictionsmentioning

confidence: 87%

Section: Development Of Predictive Correlationmentioning

confidence: 99%

Section: Validation Of Predictionsmentioning

confidence: 99%

See 1 more Smart Citation

Prediction and optimization of gene transfection and drug delivery by electroporation

Canatella

Prausnitz

2001

Gene Ther

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“…The entry of large molecules into the cell is demonstrable even when molecules are introduced into the bath up to many hours after the end of the electrical barrage [8][9][10][11][12]. Therefore, the pores must remain open for long periods.…”

Section: Is Membrane Integrity Required?mentioning

confidence: 99%

Is the Cell a Gel-and Why Does It Matter?

Pollack

2001

JJP

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It has been long recognized that the cytoplasm is a gel, even before the classic book by Frey-Wyssling [1]. Textbooks continue to emphasize the cytoplasm's gel-like consistency, and several groups have recently gone on to describe the functional relevance of the gel-sol transition [2,3]. Researchers working on actin filaments in particular have focused on the cytoplasm's gel-like behavior [4], as have some groups concerned with cellular metabolism [5]. There can be little doubt that the cytoplasm has the consistency of a gel.Yet virtually all cell biological mechanisms build on the principle of free diffusion in aqueous solution. One merely needs to examine representative mechanistic pathways to note the many diffusional steps required in proceeding from stimulus to action. These steps invariably include: ions diffusing into and out of channels; ions diffusing into and out of pumps; ions diffusing through the cytoplasm; proteins diffusing toward other proteins; and substrates diffusing toward enzymes. A cascade of multiple diffusional steps underlies nearly every intracellular process-notwithstanding the cytoplasm's character as a gel, where diffusion can be extremely slow.Surrounding this aqueous intracellular "solution" is the impermeant cell membrane, which restricts the passage of ions and other solutes to flow largely through an array of channels and pumps. Well over 100 solute-specific channels have been identified, with new ones emerging regularly. The same goes for pumps: Since ion concentrations inside and outside the cell are rarely in electrochemical equilibrium, the observed intracellular concentrations are maintained by active pumping. For pump and channel systems to operate, ions require free access.These concepts are certainly familiar to most readers of this journal. For others, the text by Alberts et al.[6] provides a detailed review of this foundational paradigm-along with the manner in which this paradigm accounts for basic cell function. Key words: phase-transition, polymer gel, cell, structured water, cooperativity, movement.Abstract: That the cell is a gel is broadly acknowledged. Textbooks begin with this assertion-and then proceed with great abandon to derive mechanisms based on free diffusion, as though the gel concept were groundless and cell was an aqueous solution. This disconnect emerges in part because the behavior of gels is not well understood, particularly among most biologists. Recently, great strides have been made in the understanding of gel behavior. It has become clear, for example, that a central mechanism in gel function is the phase-transition-a qualitative structural change prompted by a subtle change of environment, not unlike the transition from ice to water. Phase-transitions are capable of doing work. If the cell is a gel, then a logical approach to understanding cell function is to understand gel function-especially whether some role may be played by the phase-transition. Here we pursue this approach. We first consider the dichotomy of the cell as a gel and the cell...

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“…The distribution of the interested property in the population has been difficult to obtain when the experiments are done in real time. Flow cytometry has been the tool of choice to bridge the total population approach and the single cell approach by providing a large number of quantitative measurements on individual cells with high throughput (Canatella et al, 2001;Michie et al, 2000;Prausnitz et al, 1993Prausnitz et al, , 1994. Unfortunately, such measurements have to be conducted after electroporation which means that the information about the kinetics of the process is missing.…”

Section: Introductionmentioning

confidence: 99%

High-throughput and real-time study of single cell electroporation using microfluidics: Effects of medium osmolarity

Wang¹,

2006

Biotechnol. Bioeng.

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Electroporation has been widely accepted as an important tool for the delivery of exogenous molecules into cells. Previous mechanistic studies have been carried out by observing either the average behavior from a large population of cells or the response from a small number of single cells. In this study, we demonstrated a novel microfluidic method with high throughput (up to 30 Hz) for real-time studies of single cell electroporation events. Electroporation occurred when cells flowed through a section of a microfluidic channel defined by special geometry. A CCD camera was used to monitor the response of cells starting from the onset of the electroporation. We studied the swelling of Chinese hamster ovary cells and the rupture of cell membrane during electroporation using this technique. We applied buffers with different osmolarities to investigate the effects of medium osmolarity, based on results from a population of single cells. We were able to establish the distributions of the rates of swelling and membrane rupture in the cell population. We also explored establishing the correlation between the property (the cell diameter) and the behavior (the swelling rate) of single cells. Our results indicated that the processes of swelling and rupture occurred more rapidly in the hypotonic or hypertonic buffers than in the isotonic buffer. Statistical analysis did not reveal strong linear correlation between the cell size and the swelling rate. These proof-of-concept studies reveal the potential of applying microfluidics to study electroporation of a cell population at single cell level in real time with high throughput. The limitations associated with this approach were also addressed.

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Quantitative study of molecular transport due to electroporation: uptake of bovine serum albumin by erythrocyte ghosts

Cited by 59 publications

References 48 publications

Prediction and optimization of gene transfection and drug delivery by electroporation

Prediction and optimization of gene transfection and drug delivery by electroporation

Is the Cell a Gel-and Why Does It Matter?

High-throughput and real-time study of single cell electroporation using microfluidics: Effects of medium osmolarity

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