DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
This action is in response to the papers filed on 07/21/2026. Claims 18-43 are currently pending as per claims filed on 07/21/2026. Claims 24 and 31 are amended as per claims filed 07/21/2026. No claims were canceled or newly added.
Applicant's election with traverse of Group I, claims 18-31 in the reply filed on 07/21/2026 is acknowledged. Applicant timely traversed the election requirement in the reply filed on 07/21/2026.
Response to Applicants' arguments in relation to restriction requirements
Applicant’s traversal arguments are essentially that the common technical feature identified by the Examiner is a special technical feature that does make a contribution over the cited prior art, thus the 4 groups of claims relate to a single general inventive concept under PCT Rule 13.1. The Applicant essentially submits that Haufova uses tetramethoxysilane (TMOS) and (3-aminopropyl)trimethoxysilane) APTrMOS as alkoxysilane precursors in a sol-gel process to form an organo-silica layer coating a liquid marble and that the process is very different than the claimed inventive method which uses hydrophobic fumed silica. Applicant submits that the combination of Haufova and Rycheckỳ (which teaches a liquid marble coated in Polytetrafluoroethylene (PTFE)), does not teach or suggest a method of generating an organoid as set forth in claim 18. Furthermore, the Applicant submits that the Chandrashekran, when combined with the teachings of Haufova and Rycheckỳ, also do not teach or suggest the presence of methylcellulose in the cell culture medium used to prepare the liquid pearl.
These arguments have been fully considered and not found persuasive.
The Examiner notes that while Haufova uses TMOS and APTrMOS as alkoxysilane precursors, these precursors, when mixed, give rise to silicon dioxide which is formed on the surface of the alginate particle surface (Figure 4). The instant claims are “open” and require “introducing cells in suspension inside the liquid pearl”. Moreover, the liquid pearl is claimed as a product by process claim. The claims are not limited to any structure imparted by the method of making. The claims do not recite that precursors of silica cannot be used, or carboxyl groups of alginate are not present and indeed the combination of TMOS and APTrMOS and n-hexane provides a hydrophobic version of silica which can create a ahydrogel. It is further noted that generating hydrophobic fumed silica occurs via a surface chemical reaction with reactive silanes, such as TMOS and APTrMOS, thus giving basis to using TMOS and APTrMOS to form hydrophobic silica which can be coated on a liquid particle. Furthermore, while the Applicant submits that the art of Rycheckỳ pertains to powder PTFE coating on a liquid marble and the art of Haufova pertains to a liquid coating of the precursors which make the silica coating, the instant claims do not recite that the hydrophobic fumed silica must be in a liquid or powder state.
In regards to the argument pertaining to including methylcellulose in cell culture media which is used to generate liquid particles, it is noted that Chandrashekran teaches the advantages of using methylcellulose as a scaffold. Furthermore, the claims recite “introducing cells in suspension insides the liquid pearl to obtain an organoid” and the liquid pearl is generated using a solution of methylcellulose, thus upon introducing the cells in suspension, one would create a scaffold-free microenvironment with the cells in suspension which is encased by the methylcellulose shell. Thus, the combined references would indeed teach the common technical method for generating an organoid using a liquid pearl system which comprises methylcellulose.
Please note that after a final requirement for restriction, the Applicants, in addition to making any response due on the remainder of the action, may petition the Commissioner to review the requirement. Petition may be deferred until after final action on or allowance of claims to the invention elected, but must be filed not later than appeal. A petition will not be considered if reconsideration of the requirement was not requested. (See § 1.181.).
Therefore, claims 18-31 are currently under examination to which the following grounds of rejection are applicable.
Priority
The present application is a 35 U.S.C. 371 national stage filing of International
Application No. PCT/EP2022/084924 filed on 12/08/2022, which claims priority to EP 21213133.8 filed on 12/08/2021.
Filing of a certified translated copy of the EP 21213133.8 is acknowledged.
Thus, the earliest possible priority for the instant application is 12/08/2021.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 06/07/2024 was filed before the mailing date of the current office action. The submission is in compliance with the provisions of 37 CFR 1.97.
Accordingly, the information disclosure statement is being considered by the
examiner.
Claim Objections
Claim 18 is objected to because abbreviations such as “3D” should be spelled out at the first encounter in the claims. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 18 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 18 recites “(b) pipetting a volume v of a solution of methylcellulose in a cell culture medium”. It is unclear if the volume v is of the solution of methylcellulose that is added to the cell culture medium, or the volume v is the cell culture media which comprises methylcellulose. Appropriate correction is required.
Claims 19-31 inherit these deficiencies insofar as they depend on claim 18.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 18-31 are rejected under 35 U.S.C. 103 as being unpatentable over Aalders et al (Methods in Molecular Biology, 2021, pages 85-102) and further in view of Maritan et al (J of Visual Experiments, 2017, pages 1-13), and as evidenced by Wikipedia (downloaded 09/02/2026, pages 1-2).
Regarding claim 18, Aalders teaches a method of deriving cardiac organoids consisting of cocultured cardiomyocytes and cardiac fibroblasts inside a hydrophobic fumed silica powder nanoparticle (abstract, page 85). Aalders teaches the method comprises:
Dissociating cardiomyocytes and cardiac fibroblasts in a single cell suspension and mixing into different ratios in culture media
Preparing a petri dish containing a hydrophobic fumed silica powder bed
Dispensing 50 μl single culture media droplet containing 50,000 cells onto the powder bed
Gently rotating the plate in a circular motion to ensure that the powder particles completely cover the surface of the liquid drop to form the liquid marble
Culturing liquid marbles for 24 h at 37 degree C in 5% CO2 and 5% O2
Further introducing a cell suspension to the pre-made liquid marble and continue culturing (page 97, 3.12 Coalescence of Liquid Marbles to Introduce Drugs, Fresh Medium or Cells)
Hence, Aalders teaches: A method for generating an organoid, the method comprising steps of:
(1) producing or providing a liquid pearl for 3D cell culture;
(2) introducing cells in suspension inside the liquid pearl to obtain an organoid; and
(3) culturing the organoid obtained,
wherein, the liquid pearl for 3D cell culture is produced by a method comprising steps of:
(a) coating a surface of a Petri dish with hydrophobic fumed silica;
(b) pipetting a volume v of a cell culture medium and placing it on the coated surface of the Petri dish; and
(c) applying a gentle orbital shake to the Petri dish to form a liquid pearl.
Aalders does not teach pipetting a volume v of a solution of methylcellulose in a cell culture medium.
However, one of ordinary skill in the art would have considered the teachings of
Maritan as this reference is analogous prior art pertaining to use of methylcellulose in culture medium for generating spheroids (i.e. organoid).
Maritan teaches a method for forming cell spheroids with cell culture medium comprising methylcellulose. Maritan teaches that methylcellulose is non-cytotoxic, has adhesion-promoting properties of which mediates cell aggregation and minimize cell monolayer formation, is inert, contains no growth factors, and is easily removed by washing, allowing isolation of spheroids for use in a variety of applications without the presence of residual matrix (Discussion, page 12, para 1).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to modify the teachings of generating organoids using a liquid particle method as taught by Aalders to include methylcellulose in the cell culture medium since Maritan teaches that methylcellulose has advantageous properties for aggregating cells to create spheroids/organoids, is inert, and permits isolation of spheroids/organoids for subsequent use. One would be motivated to do so to more efficiently produce organoids and allow for easier succeeding isolation of the organoid from the liquid pearl. As use of methylcellulose for 3D culture and generation of liquid particles with hydrophobic fumed silica has been shown in the art, one would have a reasonable expectation of success.
Regarding claim 19, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Maritan teaches the spheroids are made in a U-bottom plate (Introduction, page 1, last para), rendering obvious (d) pipetting the liquid pearl formed in step (c) and placing it in a well of a U-bottom multiwell plate.
Regarding claims 20-22, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Maritan teaches the methylcellulose solution is at a concentration of 1-5 mg/mL in the appropriate culture medium.
Note that the concentration 1-5 mg/mL is equivalent to 0.1% to 0.5% (w/v) of methylcellulose (e.g. The weight/volume percentage expresses grams of solute per 100 mL of solution. Formula: % w/v = mg/mL ÷ 10; hence, 0.1% (w/v) = 1 mg/mL ÷ 10), thus Maritan renders obvious wherein the solution of methylcellulose in a cell culture medium comprises between 0.1 and 0.7% (w/v) of methylcellulose (claim 20), between 0.1 and 0.7% (w/v) of methylcellulose (claim 21), and 0.3% (w/v) of methylcellulose (claim 22).
Regarding claim 23, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Aalders teaches the culture media can be DMEM/F12 (page 88) or any culture media (2.6 Microbioreactor and Liquid Marble Preparation and Manipulation, page 89), rendering obvious wherein the cell culture medium is Dulbeccos Modified Eagles Medium (DMEM).
Regarding claim 24, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Aalders teaches the droplets of culture media to generate the liquid marble is 50 μl, falling within the range of 50 μl – 250 μl recited in claim 24.
Regarding claim 25 and 26, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Aalders teaches the cell suspension for liquid marble preparation is 1x106 cells/mL (page 94, 3.8 Preparation of Liquid Marbles Microbioreactors), falling within the claimed ranges 1x102 cells/mL to 1x109 cells/mL (claim 25) and range of 1x103 cells/mL to 5x108 cells/mL or of 1x104 cells/mL to 1x107 cells/mL or of 1x105 cells/mL to 5x106 cells/mL (claim 26).
Regarding claim 27, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Aalders teaches the cells in suspension are differentiated cardiomyocyte and cardiac fibroblast cells from human stem cells (page 86, para 3), rendering obvious wherein the cells in suspension are stem cells, progenitor cells or differentiated cells of human or mammal origin.
Regarding claim 28-30, the teachings of Aalders and Maritan render obvious claim 18 and 27. Moreover, Maritan teaches that spheroids generated from the methylcellulose protocol can be derived from immortalized cell lines such as SH-SY5Y, BxPC-3, and others which originate from diseased tissue (i.e. BxPC-3 is a pancreatic cancer cell line derived from 61-year old female subject; as evidenced by Wikipedia (downloaded 09/02/2026, page 1) (recited in Table 1, page 11).
Regarding claim 31, the teachings of Aalders and Maritan render obvious claim 18. Moreover, Aalders teaches drops with cell suspensions can be introduced into the liquid marble (page 97, 3.12 Coalescence of Liquid Marbles to Introduce Drugs,
Fresh Medium or Cells) and the drop size is suggested to be 10-20 μl (Note 16, page 101), falling within the range of 2 μl and 30 μl as recited in claim 31.
Conclusion
No claims are allowed.
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/JULIANA IRENE CANDELARIA/Examiner, Art Unit 1634
/MARIA G LEAVITT/Supervisory Patent Examiner, Art Unit 1634
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