Apparatuses and methods for gel molding and culture

A gel molding apparatus is adapted to be used in combination with a vessel that has a plurality of solution chambers, and includes a lid plate and a plurality of columns that project from a top surface of the lid plate. Each of the columns has a bottom side connected to the top surface of the lid plate, a top side opposite to the bottom side, and a well recessed from the top side for receiving a gel suspension and having a depth from the top side. A method for gel molding is conducted via the gel molding apparatus. A culture apparatus includes the vessel and the gel molding apparatus. A method for culture is performed through the culture apparatus.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to Taiwanese Application No. 098113647, filed Apr. 24, 2009, the disclosure of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to apparatuses and methods for gel molding and culture, more particularly to apparatuses and methods for high-throughput three-dimensional gel molding and culture.

2. Description of the Related Art

In research related to life science, culture (e.g., cell culture) is a basic laboratory technique. Cell culture mostly used in research is monolayer culture that belongs to 2-D (two-dimensional) culture. Cells show different physiology in 2-D and 3-D (three-dimensional) cell culture systems.

The 3-D cell culture system is able to establish environments that are more similar to in vivo environments, thereby inducing cell behavior that is more analogous to in vivo cell behavior. Consequently, 3-D cell culture is gradually regarded, and is widely adopted in studies related to drug screening and cell biology. In most processes of 3-D cell culture, gel is frequently utilized to entrap cells into a 3-D structure thereof. Therefore, preparation of the gel is required and includes quantification, dispensing, and molding of a gel suspension. However, conventional equipments and devices for liquid dispensing and quantification are not suitable for the gel suspension on account of viscous nature thereof. Thus, dispensing and quantification of the gel suspension may be inaccurate and may influence accuracy of experimental results.

Since high-throughput 3-D cell culture (e.g., 3-D cell culture using a commercial multi-well microplate) has been applied to numerous tasks (such as drug screening and toxin testing), a large amount of 3-D gel having small volume must be produced precisely and simultaneously. Accordingly, an apparatus and a method for efficiently achieving the aforementioned goal are in demand.

SUMMARY OF THE INVENTION

The object of the present invention is to provide apparatuses and methods for gel molding and culture in order to overcome the aforesaid drawbacks of the prior art.

According to one aspect of this invention, there is provided a gel molding apparatus that is adapted to be used in combination with a vessel having a plurality of solution chambers, and that includes a lid plate and a plurality of columns. Each of the columns projects from a top surface of the lid plate, and has a bottom side that is connected to the top surface of the lid plate, a top side that is opposite to the bottom side, and a well that is recessed from the top side for receiving a gel suspension and that has a depth from the top side. The depth of the well is relatively smaller than a height of a respective one of the columns from the top surface of the lid plate.

According to another aspect of this invention, there is provided a culture apparatus that includes a vessel and a gel molding apparatus. The vessel has a plurality of solution chambers. The gel molding apparatus includes a lid plate, and a plurality of columns that respectively have bottom sides connected to a top surface of the lid plate, top sides opposite to the bottom sides, and wells which are recessed from the top sides, respectively, for receiving a gel suspension. A depth of the wells from the topsides is relatively smaller than a height of the columns from the top surface of the lid plate. When the lid plate covers the vessel, the columns extend respectively into the solution chambers.

According to yet another aspect of this invention, there is provided a method for gel molding. The method comprises: providing a lid plate that has a plurality of columns, each of which has a well recessed from a top side thereof, and a perforated plate that has a plurality of through-holes; inserting the columns respectively into the through-holes so that the perforated plate is fitted to the lid plate; delivering a gel suspension onto the top sides of the columns and a top side of the perforated plate; and removing an excess amount of the gel suspension from the columns and the perforated plate such that the gel suspension is left in the wells.

According to still another aspect of this invention, there is provided a method for culture. The method comprises: providing a vessel that has a plurality of solution chambers, and a lid plate that has a plurality of columns having top sides respectively formed with wells; dispensing a gel suspension into the wells so that the gel suspension coagulates to form a plurality of gel modules; and inserting the columns into the solution chambers such that the lid plate covers the vessel and the gel modules contact a culture medium in the solution chambers.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Referring toFIGS. 1(a),1(b),2(a), and2(b), the preferred embodiment of a gel molding apparatus100according to the present invention is adapted to be used in combination with a vessel300(seeFIGS. 6(a) and6(b)) that has a plurality of solution chambers30(seeFIGS. 6(a) and6(b)), and includes a mold1.

The mold1has a lid plate11and a plurality of columns12that project from a top surface of the lid plate11, and may be made of a soft or rigid polymeric material. In this embodiment, the mold1is designed as a multi-well microplate lid. The columns12are spaced apart from each other so that gaps13are formed among the columns12. Each of the columns12has a bottom side121that is connected to the top surface of the lid plate11, a top side123that is opposite to the bottom side121, a surrounding wall122that interconnects the top and bottom sides123,121, and a well120that is recessed from the top side123for receiving a gel suspension and that has a depth from the top side123. The depth of the well120is relatively smaller than a height (h) of a respective one of the columns12from the top surface of the lid plate11. Precisely speaking, the height (h) of each of the columns12is equal to a distance between the top and bottom sides123,121.

Specifications for the columns12can be customized according to the demand of end-users. For instance, a specification for the columns12can be customized so as to conform to a specification for wells of a culture device. Consequently, the height of the columns12is variable. For example, for a 96-well microplate (not shown), the height of the columns12may range from 5 mm to 10 mm so that top portions of the columns12can be immersed in a liquid (e.g., a liquid culture medium, a biochemical solution, or a reagent) inside the solution chambers30of the vessel300(seeFIGS. 6(a) and6(b)).

The wells120are adapted to mold and hold gel modules that have cells therein, and hence define the shapes and specification of the gel modules, such as the thicknesses and total volumes of the gel modules. The size of the wells120can be customized when needed. If the mold1is applied to the aforementioned 96-well microplate, the wells120may be provided with diameters ranging from 3 mm to 5 mm and depths ranging from 500 μm to 2 mm. When the gel molding apparatus100is commercialized, various specifications thereof can be provided in the market.

The gel molding apparatus100further includes a perforated plate2that is made of a rigid polymeric material. Referring back toFIGS. 2(a) and2(b), the perforated plate2is fitted removably in the gaps13(seeFIG. 1(b)) among the columns12and has a plurality of through-holes20. Specifically, the columns12respectively extend through the through-holes20when the perforated plate2is fitted in the gaps13(seeFIG. 1(b)) among the columns12. Each of the through-holes20has a depth substantially equal to the height (h) of the columns12so that a top surface21of the perforated plate2is substantially flush with the surfaces of the top sides123of the columns12. The resulting flush surfaces facilitate dispensing of a gel suspension into the wells120.

According to the present invention, the preferred embodiment of a method for gel molding is described as follows. Referring toFIGS. 1(a),1(b), and5, in step601, the mold1and the perforated plate2are provided. Referring toFIGS. 2(a),2(b), and5, in step602, the columns12of the mold1are respectively inserted into the through-holes20of the perforated plate2so that the perforated plate2is fitted to the lid plate11. Referring toFIGS. 3(a),3(b), and5, in step603, a gel suspension5is delivered onto the top sides123of the columns12and the top surface21of the perforated plate2, and a scraper6is used for dispensing the gel suspension5into each of the wells120. An excess amount of the gel suspension5is removed from the top sides123of the columns12and the top surface21of the perforated plate2by dint of the scraper6such that the gel suspension5is left in the wells120. The gel suspension5contains desired biological samples such as cells, tissues, or microbes, or chemicals such as drugs. The gel suspension5may be made of a gel material for 3-D cell culture. Examples of the gel material include natural and synthetic hydrogel. Referring toFIGS. 4(a),4(b), and5, in step604, when the gel suspension5(seeFIGS. 3(a) and3(b)) coagulates to form a plurality of gel modules5′, the perforated plate2(seeFIGS. 3(a) and3(b)) is removed from the mold1. Thus, the method for gel molding is completed, and is able to efficiently and simultaneously produce a large number of the gel modules5′ that have the same specifications. The gel modules5′ are suitable for 3-D cell culture, immobilization of cells and biological molecules, drug testing and screening, toxin testing, enzyme immobilization, and the study of drug delivery.

Referring toFIGS. 6(a) and6(b), the preferred embodiment of a culture apparatus100′ according to the present invention includes the mold1and the vessel300that has a plurality of the solution chambers30. When the lid plate11of the mold1covers the vessel300, the columns12of the mold1extend respectively into the solution chambers30. It should be noted that the culture apparatus100′ could further include the perforated plate2(seeFIGS. 2(a) and2(b)) in other embodiments.

In this embodiment, the vessel300is a multi-well microplate. Each of the solution chambers30has a depth (H) that is greater than the height (h) of each of the columns12.

According to the present invention, the preferred embodiment of a method for culture is described below. Referring toFIGS. 6(a) and7, in step701, the vessel300and the mold1are provided. A culture medium4(shown inFIG. 6(b)) is disposed in the solution chambers30before the method is conducted. Referring toFIGS. 3(a),3(b),4(a),4(b), and7, in step702, the gel suspension5is delivered into the wells120so that the gel suspension5coagulates to form a plurality of the gel modules5′. The formation of the gel modules5′ is performed through steps601-604(shown inFIG. 5)of the method for gel molding. By virtue of the perforated plate2and the scraper6, the gel suspension5can be dispensed into the wells120. Referring toFIGS. 6(a),6(b), and7, in step703, the columns12are inserted into the solution chambers30such that the lid plate11covers the vessel300and the gel modules5′ contact the culture medium4in the solution chambers30. Since the gel modules5′ can be immersed in the culture medium4, the biological samples in the gel modules5′ are able to obtain nutrients in the culture medium4.

Some advantages with regard to the gel molding apparatus100, the culture apparatus100′, and the methods for gel molding and culture are as follows:

1. The mold1is compatible with a suitable commercial laboratory culture vessel (e.g., a multi-well microplate, a bioreactor, etc.), and can serve as a lid to cover the culture vessel, thereby providing a sterile environment for cell culture. In addition, the mold1has a simple structure such that a production cost thereof is low, can be easily operated, and is an efficient high-throughput 3-D culture device.

2. The 3-D gel modules5′ in the wells120of the mold1can be easily and rapidly separated from the culture medium4in the solution chambers30of the vessel300.

3. The gel modules5′ can be stained with reagent for further high-throughput analysis employing an optical screening device, such as a microplate reader.