Fluidics device allowing fluid flow between a plurality of wells

Disclosed are fluidics devices and assemblies allowing for fluid flow between a plurality of wells. The fluidics devices and assemblies that are provided mimic in vivo tissue environments by allowing for initially segregated tissue cultures that can then be linked through fluid flow to measure integrated tissue response. The devices and assemblies provide a pumpless system using surface tension, gravity, and channel geometries. By linking human tissue functional systems to better simulate in vivo feedback and response signals between the tissues, the need for testing in animals can be minimized.

TECHNICAL FIELD

This disclosure is related to a fluidics device for allowing fluid flow between a plurality of wells.

BACKGROUND

It is estimated to cost on the order of $1 B dollars to bring a drug candidate to market and the pharmaceutical industry is enhancing its chances of success by investing in human pre-clinical research. This money has driven the absorption, distribution, metabolism, elimination, and toxicology (ADMET) market in human-based products to a $5 billion dollar annual industry. The current technology for testing drug candidates is based on homogeneous culture techniques and animal models. Thus, there is an unmet need for biotool devices capable of linking human tissue functional systems to better simulate in vivo feedback and response signals between tissues and to minimize testing in animals.

Accordingly, such biotool devices and assemblies are provided in the present disclosure.

SUMMARY

Disclosed herein is a fluidics device for allowing fluid flow between a plurality of wells. The fluidics device includes a dosing well positioned upstream from a plurality of wells for containing a respective host fluid and one or more channels extending between adjacent upstream and downstream wells to define a channel fluid flow path there between, such that a dosing fluid deposited into the dosing well flows to the respective host fluid of the adjacent downstream well along the channel fluid flow path there between, and the respective host fluid subsequently flows to each adjacent downstream well along the channel fluid flow path there between.

According to one or more embodiments, the fluidics device can include a wick downstream from at least a portion of the plurality of wells. The wick is in fluid contact with the channel fluid flow path for regulating fluid flow through the plurality of wells.

According to one or more embodiments, the fluidics device can include a collection well downstream from the plurality of wells to collect the respective host fluid after having flowed through the plurality of wells. The collection well of the fluidics device can define an aperture, wherein the aperture is defined at a lower portion of a floor of the collection well.

According to one or more embodiments, the wick can be contained in the collection well such that the wick is in fluid contact with the channel fluid flow path for regulating fluid flow through the plurality of wells.

According to one or more embodiments, the surface of one or more of the plurality of wells of the fluidics device can be modified with one or both of a chemical layer or a protein layer to support a cell culture. The protein layer for supporting the cell cultures can include one or more of collagen I, collagen II, collagen III, laminin, or fibronection, or combinations thereof.

Disclosed herein is an assembly for allowing fluid flow between a plurality of wells. The assembly includes one or more fluidics devices nestably engaged and one or more reservoir trays nestably engaged on top of the fluidics device(s). The reservoir tray includes at least one chamber for containing a respective chamber fluid and an aperture defined in the chamber floor and configured such that the aperture is positioned above a dosing well of the fluidics device when in nesting engagement with the fluidics device. The floor of the chamber is angled and the aperture is defined at a lower portion of the chamber floor such that the chamber fluid flows through the aperture into the dosing well when the reservoir tray and the fluidics device are nestably engaged.

According to one or more embodiments, the assembly can include one or more reservoir trays nestably engaged underneath the fluidics device(s). The fluidics device can include a collection well downstream from the plurality of wells and the collection well defines an aperture such that fluid from the collection well flows through the aperture into the chamber of the reservoir tray nestably engaged underneath the fluidics device(s).

According to one or more embodiments, the assembly can further include a cover tray configured for nesting engagement on top of the reservoir tray nestably engaged on top of the fluidics device.

DETAILED DESCRIPTION

The presently disclosed subject matter provides fluidics devices and assemblies that in one aspect are capable of linking human functional systems to better simulate in vivo feedback and response signals between tissues and to minimize the need for testing in animal models. For example, the devices and assemblies of the presently disclosed subject matter can mimic in vivo tissue environments by allowing for initially segregated tissue cultures that can then be linked through fluid flow to measure integrated tissue response. The devices and assemblies of the present disclosure can allow for cell culture integration and media flow activated on demand. The devices and assemblies of the presently disclosed subject matter can provide a pumpless system using surface tension, gravity, and channel geometries. The devices and assemblies of the present disclosure can provide timed and tempered nutrient flow through integrated channels. The devices and assemblies of the present disclosure can provide an option to induce toxin exposure (e.g., drug exposure) at a particular cell site.

FIG. 1is a perspective view of a fluidics device100in accordance with embodiments of the present disclosure. The fluidics device100can include a dosing well110positioned upstream from a plurality of wells120for containing a respective host fluid, and one or more channels130extending between adjacent upstream and downstream wells120to define a channel fluid flow path130there between such that a dosing fluid deposited into the dosing well110flows to the respective host fluid of the adjacent downstream well120along the channel fluid flow path130there between, and the respective host fluid subsequently flows to each adjacent downstream well120along the channel fluid flow path130there between.

According to one or more embodiments, the fluidics device100can have a structure such that each adjacent downstream well120is oriented in a step-down position relative to its adjacent upstream well120. An example of a fluidics device100having this step-down well positioning structure is shown inFIG. 1.

According to one or more embodiments, the fluidics device100can include a wick140downstream from at least a portion of the plurality of wells120. The wick140is in fluid contact with the channel fluid flow path130for regulating fluid flow through the plurality of wells120. For purposes of the specification and claims, the term “wick” is meant to be used in the broadest sense to refer to a piece of material that can convey liquid by capillary action.

According to one or more embodiments, the fluidics device100can include a dosing well channel150extending from a bottom of the dosing well110to the channel fluid flow path130such that the dosing fluid flows to the respective host fluid of the adjacent downstream well120through the dosing well channel150and along the channel fluid flow path130. A side of the dosing well110can define an angle of greater than 90° extending from a bottom of the dosing well110up to the channel fluid flow path130of the adjacent well120. According to one or more embodiments, the fluidics device100can include a collection well170downstream from the plurality of wells120to collect the respective host fluid after having flowed through the plurality of wells120. The collection well170of the fluidics device100can include a floor that defines a divot180, wherein the floor is angled such that the divot180is defined at a lower portion of the floor. In certain embodiments according to the present disclosure as described herein below, the lower portion of the floor of the collection well170can define an aperture as an alternative to the divot180. In another example, the divot180can be converted to an aperture for use of the fluidics device100in an assembly as described herein below.

In accordance with embodiments of the present disclosure, the collection well170of fluidics device100can include one or more collection well channels210extending from the channel fluid flow path130to a bottom of the collection well170such that the respective host fluid of the adjacent upstream well120flows along the channel fluid flow path130and through the collection well channel210into the collection well170. The collection well channel210can have a width ranging from about 10 to 3500 microns and a depth ranging from about 10 to 3500 microns. The collection well170can define a ramp extending from a bottom of the collection well170up to the channel fluid flow path130of the adjacent upstream well120. The ramp can include 1, 2, 3, or 4 of the collection well channels210that are contiguous with the ramp.

FIG. 2is a perspective view of the fluidics device100in accordance with embodiments of the present disclosure.FIG. 2illustrates that the fluidics device100can include a dosing well channel cover160configured to enclose the dosing well channel150. An example of the dosing well channel cover160is shown inFIG. 2where each of the 8 dosing wells (A-H) are covered with the dosing well channel cover160.FIG. 2also illustrates that the fluidics device100can include a channel cover230configured for engagement on top of the one or more channels130extending between the adjacent wells120to enclose the channels130.

The wick of the present disclosure can define any shape that is suitable for being in fluid contact with the channel fluid flow path130and for regulating fluid flow through the plurality of wells120. For example, the wick of the presently disclosed subject matter can be any absorbent material. The wick can regulate fluid flow through the plurality of wells120at a rate ranging from 0.0007 ml/min to 30 ml/min. In one example, the respective host fluid after having flowed through each of the plurality of wells120and onto the wick can evaporate off the wick.

FIG. 2illustrates two examples of the wick (i.e wick140and wick142) at separate positions downstream from a portion of the plurality of wells120.FIGS. 3A-3Cillustrate the wick in accordance with one or more embodiments of the present disclosure.FIG. 3Aillustrates an example of the wick142having a cylindrical shape.FIG. 3Cillustrates an example of the wick140defining a generally flat shape. The wick defining a generally flat shape can define a gap such that only a portion of an edge of the wick is in fluid contact with the channel fluid flow path130.FIG. 3Cillustrates an example of the wick144defining a gap190.

The wick142defining a cylindrical shape is illustrated inFIG. 2andFIG. 3A. The wick of the present disclosure can be positioned anywhere downstream from at least a portion of the plurality of wells120. For example, the wick142defining a cylindrical shape is shown contained in well120in row eight of the fluidics device100inFIG. 2such that the wick142is in fluid contact with the channel fluid flow path130for regulating fluid flow through the plurality of upstream wells120.

The wick can define a generally flat shape. According to one or more embodiments, the wick140or144defining a generally flat shape can be contained in the collection well170such that the wick140or144is in fluid contact with the channel fluid flow path130for regulating fluid flow through the plurality of wells120. The wick defining a generally flat shape can be carried by a shoulder defined by the collection well170such that the wick does not contact a bottom surface of the collection well170. An example of the wick140defining a generally flat shape and carried by a shoulder defined by the collection well170is illustrated inFIG. 2and inFIG. 3B. The wick defining a generally flat shape can be carried by one or more posts defined by the collection well170such that the wick does not contact a bottom surface of the collection well170. In one embodiment, the wick can define a generally flat shape and can be carried by six of the posts defined by the collection well170such that the wick does not contact a bottom surface of the collection well170.

FIG. 4illustrates the dosing well channel cover160separate from the fluidics device100.

FIG. 5illustrates a top view of the fluidics device ofFIG. 1showing an enlarged view of the dosing well110, dosing well channel150, adjacent downstream well120, and the one or more channels130extending there between in accordance with embodiments of the present disclosure. The dosing well channel150can have a width ranging from about 10 to 3500 microns and a depth ranging from about 10 to 3500 microns. The dosing well channel150can include 2, 3, or 4 channels contiguous with the dosing well channel150and each of the channels can have a width ranging from about 200 to 1500 microns and a depth ranging from about 10 to 1500 microns.

The one or more channels130extending between adjacent upstream and downstream wells120of the fluidics device100can have a width ranging from 10 to 3500 microns and a depth of 10 to 1500 microns. An example of a fluidics device100having a single channel130is shown inFIG. 5. The channel130can define a triangular-shape that extends between each of the adjacent wells120. The triangular-shape channel130can be positioned such that the triangular shape generally converges at each adjacent downstream well120. An example of a fluidics device100having the triangular-shape channel130positioned such that the triangular shape generally converges at each adjacent downstream well120is shown inFIG. 5.

The fluidics device100can have 2, 3, or 4 channels130and each of the channels130can have a width ranging from 200 to 750 microns and a depth ranging from 10 to 1500 microns. The fluidics device100can include 2, 3, or 4 microchannels200that are contiguous with the channel130and each of the microchannels200can have a width ranging from 200 to 750 microns and a depth ranging from 10 to 1500 microns. An example of a fluidics device100having 3 microchannels200that are contiguous with the triangular-shape channel130is shown inFIG. 5.

The channel cover230can include 1 or more projections extending from the channel cover230such that when the channel cover230is engaged on top of the channels130of the fluidics device100the channel cover230defines 2 or more microchannels200contiguous with the channel130. For example, the channel cover230can have two projections such that when the channel cover230is engaged on top of the channel130of the fluidics device100the channel cover230defines 3 microchannels200contiguous with the channel130. In one embodiment, each of the microchannels200defined by the channel cover230can have a width ranging from 200 to 750 microns and a depth ranging from 10 to 1500 microns.

The bottom surface of each of the channels130, the dosing well channel150, the microchannels200, and the collection well channels210can define different shapes. For example, the channels130, the dosing well channel150, the microchannels200, and the collection well channels210can define an arcuate bottom surface or a generally flat bottom surface.

According to one or more embodiments, the fluidics device100can have a structure where the plurality of wells120are aligned in a row. The fluidics device100can have 12 wells in a respective row and a total of 8 rows. An example of a fluidics device100having this structure is shown inFIGS. 1, 2, and 8. The fluidics device100can have 3 wells in a row and a total of 2 rows. The fluidics device100can have 6 wells in a row and a total of 4 rows. The fluidics device100can have 8 wells in a row and a total of 6 rows. The fluidics device100can have 12 wells in a row and a total of 8 rows. The fluidics device100can have 24 wells in a row and a total of 16 rows. The fluidics device100can have 48 wells in a row and a total of 32 rows.

According to one or more embodiments, the fluidics device100can have a structure where the plurality of wells120for containing a respective host fluid are oriented in a configuration such that each downstream well120is positioned lower relative to each adjacent upstream well120and the dosing well110is upstream from the plurality of wells120and in fluid communication therewith.

The fluidics device100of the presently disclosed subject matter can be employed for any use requiring the tempered flow of fluid between a plurality of wells. According to one or more embodiments, a method for employing the fluidics device100includes adding a dosing fluid to the dosing well110and adding the respective host fluid to the plurality of wells120such that the fluid is in fluid contact with the channel fluid flow path130, whereby the dosing fluid flows to each of the respective host fluids in the plurality of wells120in a tempered manner. The method can include removing an aliquot of the respective host fluid from the wells120at one or more time periods to measure the effect of the dosing fluid being tempered through the plurality of wells120over time.

The dosing fluid can include, for example, but is not limited to a drug, a legal or illegal drug, a toxin, an agent of warfare, a fragrance, a food spice, an oil, a gas, a metabolite, a compound, a hormone, a solution, a solute, a composite, a nutrient media, differentiation media, or a growth media, and combinations thereof. The plurality of wells120can contain a respective cell culture whereby an effect of the tempered exposure to the dosing fluid on the cells can be measured. The effect of the tempered exposure to the dosing fluid on the cell cultures to be measured can be one or more of pharmacokinetics, drug metabolism, toxicity, pre-clinical pharmaceutical studies, cell response, cell receptor response, cell feedback signals, cell growth, cell death, cell differentiation, or cell regeneration, and combinations thereof. The respective cell culture can be, for example, a stem cell culture or a progenitor cell culture.

According to one or more embodiments, the plurality of wells120of the fluidics device100can contain a respective cell culture, and a method for employing the fluidics device100containing the respective cell cultures includes adding a dosing fluid to the dosing well110, adding the desired respective host fluid to the wells120such that the fluid is in fluid contact with the channel fluid flow path130. Subsequently, the dosing fluid flows to each of the respective host fluids in the plurality of wells120in a tempered manner. The method can further include removing an aliquot of the respective host fluid from the wells120at one or more time periods to measure the effect of the dosing fluid on the cells.

The fluidics device100can be made of any material that is suitable for use in fluid transfer between the plurality of wells120. The type of material chosen can depend on the desired use of the fluidics device100. For example, the user of the fluidics device100can choose the material based on the dosing well fluid that will be used and the expected interaction of the dosing well fluid with the material. Thus, the fluidics device100can be made of any suitable material including, for example, a polymer, a synthetic polymer, a TOPAS® COC polymer, a biodegradable polymer, a plastic, a biodegradable plastic, a thermoplastic, a polystyrene, a polyethylene, a polypropylene, a polyvinyl chloride, a polytetrafluoroethylene, a silicone, a glass, a PYREX, or a borosilicate, or combinations thereof. In addition, the dosing well channel cover160, the channel cover230, and the wick140,142,144may each be made from the same materials as the fluidics device100. In one example, a user may wish to have each of the fluidics device100, the dosing well channel cover160, the channel cover230, and the wick140,142,144made from the same material such that the interaction of the dosing well fluid with the material does not vary.

According to one or more embodiments, the surface of one or more of the plurality of wells120of the fluidics device100can be modified with one or both of a chemical layer or a protein layer to support a cell culture. The protein layer for supporting the cell cultures can include one or more of collagen I, collagen II, collagen III, laminin, or fibronection, or combinations thereof.

According to one or more embodiments of the presently disclosed subject matter, an assembly is provided for allowing fluid flow between the plurality of wells120of the fluidics device100. The assembly can include the fluidics device100and a reservoir tray250configured for nesting engagement on top of the fluidics device100.FIG. 6shows a perspective view of the reservoir tray in accordance with embodiments of the present disclosure.FIG. 7shows a bottom view of the reservoir tray in accordance with embodiments of the present disclosure. According to one or more embodiments, an assembly is provided that includes the fluidics device100, the reservoir tray250, and a cover tray260configured for nesting engagement on top of the reservoir tray250or the fluidics device100.FIG. 8shows an exploded perspective view of the fluidics device100as part of an assembly including the reservoir tray nestably engaged on top of the fluidics device100and the cover tray260nestably engaged on top of the reservoir tray in accordance with embodiments of the present disclosure.

According to one or more embodiments of the presently disclosed subject matter, an assembly is provided for allowing fluid flow between the plurality of wells120of the fluidics device100, the assembly including the fluidics device100and the reservoir tray250configured for nesting engagement on top of the fluidics device100. Turning to FIG.'s6and7, the reservoir tray250can include at least one chamber270for containing a respective chamber fluid and an aperture280defined in the chamber floor and configured such that the aperture280is positioned above the dosing well110of the fluidics device100when in nesting engagement with the fluidics device100. The floor of the chamber270can be angled and the aperture280can be defined at a lower portion of the chamber floor such that the chamber fluid flows through the aperture280into the dosing well110when the reservoir tray250and the fluidics device100are nestably engaged. When nestably engaged, the reservoir tray250can be positioned just above the fluidics device100and the respective chamber fluid flows from each chamber270of the reservoir tray250through each aperture280and into each dosing well110of the fluidics device100.

According to one or more embodiments, the assembly can further include the cover tray260configured for nesting engagement on top of the reservoir tray250of the fluidics device100. According to one or more embodiments, the assembly can include one or more additional reservoir trays250configured for nesting engagement on top of the fluidics device100.

According to one or more embodiments of the presently disclosed subject matter, the assembly can further include a second reservoir tray250configured for nesting engagement underneath the fluidics device100.FIG. 9shows an exploded side view of this assembly including the second reservoir tray250in accordance with embodiments of the present disclosure. For this assembly, the fluidics device100can include the collection well170that is downstream from the plurality of wells120and the collection well170can define an aperture such that when the second reservoir tray250is in nesting engagement underneath the fluidics device100, fluid from the collection well170of the fluidics device100flows through the aperture into the chamber270of the second reservoir tray250. Referring toFIG. 9, the fluid can flow from the reservoir tray250nestably engaged on top of the fluidics device100from right to left through the aperture280of the reservoir tray250into the dosing well110of the fluidics device100. The fluid can flow from the dosing well110from left to right through the aperture of the collection well170of the fluidics device100into the chamber270of the reservoir tray250nestably engaged underneath the fluidics device100. The fluid can then flow in the second reservoir tray250from right to left.

According to one or more embodiments, the assembly can include one or more additional reservoir trays250configured for nesting engagement underneath the fluidics device100.

According to one or more embodiments of the presently disclosed subject matter, the assembly can include a second fluidics device100configured for nesting engagement underneath the fluidics device100. In this assembly, the fluidics device100can include the collection well170downstream from the plurality of wells120and the collection well170can define an aperture such that fluid from the collection well170flows through the aperture into the dosing well110of the second fluidics device100underneath when the fluidics devices100are nestably engaged.

According to one or more embodiments of the presently disclosed subject matter, the assembly can include one or more additional fluidics devices100configured for nesting engagement underneath the second fluidics device100. The additional fluidics devices100can each include the collection well170downstream from the plurality of wells120and each collection well170can define an aperture such that fluid from the collection well170flows through the aperture into the dosing well110of the additional fluidics device100positioned underneath when the multiple fluidics devices100are nestably engaged.FIG. 10shows an exploded side view of this assembly including a total of three fluidics devices100nestably engaged, reservoir trays250engaged on top of and underneath the three fluidics devices100, and cover tray260engaged on top of the top reservoir tray250in accordance with embodiments of the present disclosure.

According to one or more embodiments of the presently disclosed subject matter, a method is provided for employing an assembly including one or more nestably engaged fluidics devices100and one or more reservoir trays250nestably engaged on top of and/or underneath the fluidics devices100as exemplified inFIGS. 8-10. The method can include adding a dosing fluid to the dosing well120and adding a respective host fluid to the plurality of wells120of the fluidics device(s)100such that the fluid is in fluid contact with the channel fluid flow path130, whereby the dosing fluid flows to each of the respective host fluids in the plurality of wells120in a tempered manner. The method includes positioning the reservoir tray250above the fluidics device(s)100such that the reservoir tray250and the fluidics device(s)100are in nesting engagement, and adding the respective chamber fluid to the respective chamber270of the reservoir tray250, whereby the respective chamber fluid flows into the dosing well110of the fluidics device100that is nestably engaged underneath the reservoir tray250. In this manner, a larger supply of dosing fluid than can be contained by the dosing well110alone can be provided at a tempered rate to the one or more fluidics devices100that are nestably engaged underneath the reservoir tray250.

According to one or more embodiments, the dosing fluid can include one or more of a drug, a legal or illegal drug, a toxin, an agent of warfare, a fragrance, a food spice, an oil, a gas, a metabolite, a compound, a hormone, a solution, a solute, a composite, a nutrient media, a differentiation media, or a growth media.

According to one or more embodiments, the plurality of wells120of the fluidics device100can contain a respective cell culture, whereby an effect of the tempered exposure to the dosing fluid on the cells can be measured. The effect of the tempered exposure to the dosing fluid on the cell cultures to be measured can be one or more of pharmacokinetics, drug metabolism, toxicity, pre-clinical pharmaceutical studies, cell response, cell receptor response, cell feedback signals, cell growth, cell death, cell differentiation, or cell regeneration. The respective cell culture can be a stem cell culture or a progenitor cell culture.

According to one or more embodiments, the method for employing the assembly can further include removing an aliquot of the respective host fluid from one or more of the plurality of wells120at one or more time periods to measure an effect of the dosing fluid from having been tempered through the plurality of wells120. The plurality of wells120can contain a respective cell culture, and the method can include removing an aliquot of the respective host fluid from one or more of the plurality of wells120at one or more time periods to measure an effect of the dosing fluid on the cells.