Embryonic development in the mouse is enhanced via microchannel culture

Raty, S.; Walters, Eric M.; Davis, John; Zeringue, Henry C.; Beebe, David J.; Rodriguez-Zas, Sandra L.; Wheeler, Matthew B.

doi:10.1039/b316437c

Cited by 153 publications

(100 citation statements)

References 28 publications

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“…Microchannels have been used for several steps in the in vitro production of embryos typically either matching or improving the performance of previous methods. 1,2 Insect cell cultures as well have shown very different dimension dependent growth kinetics in microscale cultures as compared to macroscale flask cultures. 3 However, much of the previous culture work in microfluidics has focused on non-adherent (e.g., embryos, insect cells) or 2-D adherent cell culture in the absence of extracellular matrix.…”

Section: Introductionmentioning

confidence: 99%

Gelatin based microfluidic devices for cell culture

Paguirigan¹,

Beebe²

2006

Lab Chip

145

139

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We have developed a technique for fabricating microfluidic devices from gelatin using a natural crosslinking process. Gelatin, crosslinked with the naturally occuring enzyme transglutaminase is molded to produce microchannels suitable for adherent cell culture and analysis. The autofluorescence of the material was shown to be minimal and within the range of typical background, ensuring utility with analyses using fluorescent dyes and labels would not be affected. Also, normal murine mammary epithelial cells were successfully cultured in the microchannels. The morphology of these adherent epithelial cells was shown to be significantly different for cells grown on rigid tissue culture plastic in either macro-or microscale cultures (even in the presence of a surface coating of gelatin) than those grown on the flexible crosslinked gelatin microchannels. Using these devices, the effects of both the extracellular matrix and soluble factors on cellular behavior and differentiation can be studied in microenvironments that more closely mimic the in vivo environment. IntroductionWhile microfluidics has shown considerable promise as a tool for studying cell biology, the potential for microfluidics to create more in vivo-like in vitro environments (or, in visio environments) is still largely untapped. It is becoming clear that the scale of the microenvironment provided by microchannels is an important biological parameter. Microchannels have been used for several steps in the in vitro production of embryos typically either matching or improving the performance of previous methods. 1,2 Insect cell cultures as well have shown very different dimension dependent growth kinetics in microscale cultures as compared to macroscale flask cultures. 3 However, much of the previous culture work in microfluidics has focused on non-adherent (e.g., embryos, insect cells) or 2-D adherent cell culture in the absence of extracellular matrix. While the effects of the extracellular matrix are of critical importance to in vivo tissue function, 4,5 the integration of extracellular matrix (ECM) materials into microfluidic systems is just emerging. 6,7 We have fabricated microdevices which can allow for control over the soluble microenvironment, while also providing a more in vivo-like substrate (flexible protein gels) as a step towards an in visio environment for cellular analysis. We present a fabrication protocol to produce enzymatically crosslinked gelatin microdevices composed of natural components, that are sterile and suitable for cell culture. Gelatin is a derivative of collagen, one of the most common extracellular matrix proteins. These devices were tested for their fluorescence properties to ensure that any autofluorescence of the material itself will not limit its utility while using fluorescent dyes for cellular analysis. Also, we have explored the potential for mammary epithelial cell culture in these devices and compared the cell morphology at multiple time points between macro-and microscale cultures on traditional rigid surfaces an...

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Section: Introductionmentioning

confidence: 99%

Gelatin based microfluidic devices for cell culture

Paguirigan¹,

Beebe²

2006

Lab Chip

145

139

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“…The use of HEK in toxicity assays (Chou et al 2002;Didier et al 1999;Monteiro-Riviere et al 2005;Rouse et al 2006;Shvedova et al 2003) makes this cell type an attractive candidate for use in microfluidic high throughput toxicity assays. Cell culture in microfluidic systems has been demonstrated by others Delamarche et al 2005;Figallo et al 2007;Flaim et al 2005;Folch and Toner 2000;Gu et al 2004;Hui and Bhatia 2007;Hung et al 2005a, Hung et al b;Kane et al 2006;Khademhosseini et al 2004;Kim et al 2006;Leclerc et al 2006a;b;Lee et al 2006;Prokop et al 2004;Raty et al 2004;Rhee et al 2005;Sin et al 2004;Song et al 2005;Toh et al 2007;Tourovskaia et al 2005;Walker et al 2002) but keratinocytes have not yet been studied at the microscale.…”

Section: Introductionmentioning

confidence: 99%

Characterization of microfluidic human epidermal keratinocyte culture

2008

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Human epidermal keratinocytes (HEK) are skin cells of primary importance in maintaining the body's defensive barrier and are used in vitro to assess the irritation potential and toxicity of chemical compounds. Microfluidic systems hold promise for high throughput irritant and toxicity assays, but HEK growth kinetics have yet to be characterized within microscale culture chambers. This research demonstrates HEK patterning on microscale patches of Type I collagen within microfluidic channels and maintenance of these cells under constant medium perfusion for 72 h. HEK were shown to maintain 93.0%-99.6% viability at 72 h under medium perfusion ranging from 0.025-0.4 ll min -1 . HEK maintained this viability while *100% confluent-a level not possible in 96 well plates. Microscale HEK cultures offer the ability to precisely examine the morphology, behavior and viability of individual cells which may open the door to new discoveries in toxicological screening methods and wound healing techniques.

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“…An alternate approach that permits a commercial means of utilizing microwells in conjunction with existing dishware in the lab involves using microwell-inserts consisting of several rows of tiny culture wells composed of PDMS [38]. Within PDMS and glass-bottomed microchannels under static media conditions, 2-cell mouse embryos can be cultured to the blastocyst stage [65,66]. However, researchers found that culture in microchannels resulted in significantly greater blastocyst formation and hatched blastocyst development at 72 h and 96 h, respectively.…”

Section: Embryo Culture Systems Using Pdms Chambers or Channelsmentioning

confidence: 99%

“…Within the last decade, studies using PDMS microfluidic channels or funnels have suggested novel solutions for oocyte manipulation, sperm sorting, and embryo culture . Microfluidic systems that mimic oviductal structures and functions for use in RT are divided into those used for sperm motility control or monitoring [50][51][52][53][54][55][56][57][58][59][60], regulation of chemical gradients for in vitro fertilization and embryo culture [61][62][63][64][65][66][67][68][69], and applying mechanical stimuli to the developing embryo [70][71][72][73][74]. Although the unique characteristics of elastomers have been exploited for microstructure fabrication and/or micro-pumping, mechanical deformation of the elastomer membrane has not been used to produce new devices for RT.…”

Section: Microfluidic Channels To Handle Sperm and Embryosmentioning

confidence: 99%