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/Restriction
Applicant’s election of Group I, claims 1-6, drawn to a method for providing structural support to EBs or a method for producing microglia-like cells, in the reply filed on 08/15/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.03(a)).
Claims 7-16 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim.
Claim Status
Claims 1-16 are pending.
Claims 7-16 are withdrawn.
Claims 1-6 are considered on the merits.
Priority
This application is a 371 of PCT/EP2021/056037 (filed on 03/10/2021), which claims benefit from foreign application EP20162230.5 (filed on 03/10/2020). The priority claim of the instant application has been granted and the earliest benefit date is 03/10/2020 from the application EP20162230.5.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 09/01/2022 and 06/04/2025 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner. The corresponding signed and initialed PTO forms 1449 have been mailed with this action.
Claim Objections
Claims 1 and 6 are objected to because of the following informalities:
Claim 1 (c) recites the phrase “cell-culture medium”. It is recommended to change to “a cell-culture medium”.
Claim 6 (ii) recites continuing cultivation in “cell-culture medium”. If this medium is the same medium as recited in claim 1 (c), it is recommended to change to “the cell-culture medium”.
Furthermore, claim 6 (ii) recites “microglial-like cells”. It is recommended to change to “microglia-like cells” to be consistent with claim 6 (iii) and specification.
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.
Claims 1-6 are 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 1 (b) recites the limitation “the microcarrier is porous”. There is insufficient antecedent basis for this limitation because step (b) only recites “a macrocarrier”. It is recommended to change the limitation to “the macrocarrier is porous”. Claims 2-6 are rejected as being dependent from claim 1 but not resolving the ambiguity.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claims 1-2 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Nestor et al., (Stem Cell Research. 2013; 10: 454-463. Cited in IDS 09/01/2022).
With respect to claim 1, Nestor teaches a method of culturing embryoid bodies on a mesh insert (see e.g., abstract), and teaches the mesh insert provides a physical support for the tissue to spread out and grow in a non-spherical manner (see p. 462, left col, last para), thus teaches a method for providing structure support to embryoid bodies for adherence and outgrowth in preamble of claim 1.
In regard to step (a) providing embryoid bodies, Nestor teaches iPS cells are plated in a 96-well V-bottom plate at a density of 9000 cells per well and are cultured for 14 days to form embryoid bodies (p. 455, left col, section “Material/methods”, para 2, also see Fig 1A leftmost panel for embryoids formed in the well).
In regard to step (b) seeding the embryoid bodies onto a porous macrocarrier, Nestor teaches at day 14, the embryoid bodies are transferred by pipetting with wide-orifice tips onto Millipore mesh inserts (MI) (0.4 µM pore size) inserted into 6-well plates and the embryoid bodies rest on the surface of the Millipore insert and are cultured until day 30 (p. 455, left col, section “Material/methods”, para 2, also see Fig 1A for transferring, resting and culturing embryoid bodies on the insert), thus teaches seeding thereby obtaining a porous macrocarrier (i.e., the Millipore mesh insert) with adherent embryoid bodies (embryoid bodies cultured on the insert for at least 16 days).
In regard to step (c) culturing the macrocarrier with the adherent embryoid bodies in a cell culture medium, as stated supra, Nestor teaches the embryoid bodies are cultured on the surface of the mesh insert for at least 16 days in a cell culture medium (day 14 – day 30, see p. 455, left col, section “Material/methods”, para 2).
With respect to claim 2 (v) directed to the size of the macrocarrier being at least 0.1 cm in one dimension, Nestor teaches multiple embryoid bodies are seeded on one mesh insert (see e.g., Fig 1A for 4 embryoid bodies separated apart at Day 14 with a distance of at least 3 diameters of embryoid bodies in between) and teaches the size of the embryoid bodies is at least 350 micrometer in one dimension at day 15 (see e.g., Fig 1E leftmost panel “Day 15” in which the scale bar is 50 µm and the diagonal length of the embryoid body is at least 7 times of the scale bar, i.e., 50 µm x 7 = 350 µm). Thus, the mesh insert shown in Fig 1A panel “Day 14” is at least 0.1 cm in one dimension (e.g., the mesh insert in one dimension has 2 embryoid bodies with a distance of at least 3 diameters of embryoid bodies in between, i.e., (2+3) x 350 µm = 1750 µm = 0.175 cm).
Accordingly, Nestor anticipates instant claims 1-2.
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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-3 are rejected under 35 U.S.C. 103 as being unpatentable over Nestor et al., (Stem Cell Research. 2013; 10: 454-463. Cited in IDS 09/01/2022).
Claims 1-2 are anticipated by Nestor as stated supra. Thus, Nestor makes obvious claims 1-2.
With respect to claim 3 directed to the seeding density of embryoid bodies on the macrocarrier, as stated supra, Nestor teaches multiple embryoid bodies are seeded on one mesh insert (see e.g., Fig 1A for 4 embryoid bodies seeded at Day 14). For the sake of estimating the area of the mesh insert, Nestor’s Fig 1B is used as a schematic diagram drawn in scale. The diameter of the insert is about 2/3 of the diameter of the well in a 6-well plate, which is known to have a diameter of about 3.6 cm, thus the diameter of the insert is about 2.4 cm and the area of the insert is about 4.5 cm2, and thus the seeding density of the embryoid bodies on the insert shown in Fig 1A is about 4 / 4.5 cm2 = 0.9 per cm2, within the claimed range of about 1 embryoid body per cm2.
Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method for providing structural support to embryoid bodies by seeding them onto a macrocarrier disclosed by Nestor, by seeding the embryoid bodies at a density within the claimed range as suggested by Nestor with a reasonable expectation of success. Since Nestor suggests a seeding density within the claimed range as estimated from Fig 1B, one of ordinary skill in the art would have had a reason to seed the embryoid bodies at the claimed density in order to provide suitable support for the embryoid bodies to spread out and grow on the macrocarrier (see p. 462, left col, last para).
Furthermore, MPEP states “generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical” and “where the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation”. See MPEP 2144.05(II)(A). In the instant case, Nestor teaches multiple embryoid bodies can be seeded onto the macrocarrier to provide physical support for the embryoid bodies to spread out and grow (see p. 462, left col, last para). Therefore, it would have been obvious for one ordinary skill in the art to apply the claimed seeding density because they are the results of “routine optimization”.
Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary.
Claims 1-2 and 4-5 are rejected under 35 U.S.C. 103 as being unpatentable over Nestor et al., (Stem Cell Research. 2013; 10: 454-463. Cited in IDS 09/01/2022) in view of Mandalam et al., (WO 2007/002086 A2. Cited in IDS 09/01/2022).
Claims 1-2 are anticipated by Nestor as stated supra. Thus, Nestor makes obvious claims 1-2.
With respect to claim 4 directed to the macrocarrier floating freely in the cell culture medium, and claim 5 directed to the macrocarrier being subject to dynamic movement during culturing, Nestor teaches the embryoid bodies rest on the surface of the mesh insert with the medium underneath (p. 455, left col, “Material/methods”, para 2, and see Fig 1B diagram), indicating the macrocarrier floats in cell culture medium.
However, Nestor does not specifically teach the macrocarrier floats freely in the medium in claim 4, or is subject to dynamic movement during culturing in claim 5.
Mandalam teaches a method of suspension culture of embryonic stem cells (see e.g., abstract). Mandalam teaches the suspension culture may contain particulate carriers that create surfaces within the suspension, but still provide the benefits of culturing the cells in a three-dimensional space, and one type of such carriers is disk-shaped culture plastic, such as the Fibra-cel Disks (p. 9, lines 15-16 and 20-21). Mandalam teaches the suspension culture may be on a shaker for long-term culture (see Example 4 in page 17 and Fig 7). Thus, Mandalam suggests a method of suspension culture for stem cells using a macrocarrier on a shaker, thus suggests the macrocarrier floats freely in the cell culture medium (i.e., in suspension culture, related to claim 4), and the macrocarrier is subject to dynamic movement during culturing (i.e., on a shaker, related to claim 5).
Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method for providing structural support to embryoid bodies by seeding them onto a macrocarrier with the medium underneath disclosed by Nestor, by combining suspension culture on a shaker such that the macrocarrier floats freely in the cell culture medium and is subject to dynamic movement during culturing as suggested by Mandalam with a reasonable expectation of success. Since Mandalam teaches the suspension culture maximizes the production capacity of the culture environment and allows for bulk proliferation in a more cost-effective manner, which facilitates commercial production of important products for use in human therapy (e.g., abstract) and the suspension culture on a shaker is suitable for long-term culture (e.g., Example 4 showing culturing over two months), one of ordinary skill in the art would have had a reason to combine suspension culture on a shaker in the method of Nestor in order to maximize the production capacity and to enable long-term culture of embryoid bodies.
Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary.
Claims 1-2 and 6 are rejected under 35 U.S.C. 103 as being unpatentable over Haenseler et al., (Stem Cell Reports. 2017; 8: 1727-1742) in view of Nestor et al., (Stem Cell Research. 2013; 10: 454-463. Cited in IDS 09/01/2022).
With respect to claim 6, Haenseler teaches a method for producing microglial precursor cells and microglia cells (see e.g., abstract and Fig 1C), thus teaches the preamble of claim 6.
In regard to claim 6 (i) referring to the steps of claim 1, Haenseler teaches defined-size embryoid bodies (EBs) are formed using Aggrewells (p. 1728, right col, “Results” para 1, see Fig 1C), thus teaches providing embryoid bodies in claim 1 (a).
However, although Haenseler teaches the embryoid bodies are plated into large-format flasks (i.e., a macrocarrier) with medium and most EBs adhere (p. 1728, right col, “Results” para 1), thus teaches seeding the EBs onto a macrocarrier thereby obtaining a macrocarrier with adherent EBs in claim 1 (b), Haenseler is silent on the macrocarrier being porous in step (b) of claim 1 or its size in claim 2.
Nestor teaches a method of culturing embryoid bodies on a mesh insert (see e.g., abstract), and teaches the mesh insert provides a physical support for the tissue to spread out and grow in a non-spherical manner (see p. 462, left col, last para), thus teaches a method for providing structure support to embryoid bodies for adherence and outgrowth in preamble of claim 1. In regard to claim 1 (b) seeding the embryoid bodies onto a porous macrocarrier, Nestor teaches the embryoid bodies are transferred by pipetting with wide-orifice tips onto Millipore mesh inserts (MI) (0.4 µM pore size) inserted into 6-well plates and the embryoid bodies rest on the surface of the Millipore insert and are cultured until day 30 (p. 455, left col, section “Material/methods”, para 2, also see Fig 1A for transferring, resting and culturing embryoid bodies on the insert), thus teaches seeding the embryoid bodies onto a porous macrocarrier (i.e., the Millipore mesh insert) thereby obtaining a porous macrocarrier with adherent embryoid bodies in claim 1 (b). Nestor teaches the size of the mesh insert shown in Fig 1A panel “Day 14” is at least 0.1 cm in one dimension (e.g., Fig 1A shows the mesh insert in one dimension has 2 embryoid bodies with a distance of at least 3 diameters of embryoid bodies in between (i.e., at least 5 times the diameter of EBs), and the embryoid bodies have a diameter of at least 350 µm, see e.g., Fig 1E leftmost panel “Day 15” compared to the scale bar), thus teaches claim 2.
Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method for producing microglial precursor cells and microglia cells through embryoid bodies disclosed by Haenseler, by substituting the culture macrocarrier with a porous mesh insert with the claimed size as suggested by Nestor with a reasonable expectation of success. Since Nestor teaches the porous mesh insert provides a physical support for the EBs to spread out and grow in a non-spherical manner with greatly enhanced cell viability (p. 462, left col, last para), and this technique has the advantage of combination of imaging and functional study (p. 462, right col, para 1), one of ordinary skill in the art would have had a reason to substitute with the porous macrocarrier (i.e., the mesh insert) with the claimed size as suggested by Nestor in the method of Haenseler in order to facilitate EB spreading out and growing with greatly enhanced cell viability and to take advantage of combination of imaging and functional study (p. 462, left col, last para and right col, para 1).
In regard to claim 1 (c), Haenseler teaches culturing the adherent EBs in cell-culture medium (p. 1728, right col, “Results” para 1), and Nestor teaches the EBs are cultured on the surface of the mesh insert for at least 16 days in a cell culture medium (day 14 – day 30, see p. 455, left col, section “Material/methods”, para 2).
In regard to claim 6 (ii), Haenseler teaches continuing cultivation of the adherent embryoid bodies in cell culture medium with M-CSF and IL-3 (see Fig 1C) to differentiate into embryonic macrophage precursors (see Fig 1C “pMacpre”, which stands for PSC derived macrophage/microglia precursors) and further differentiate into microglia (“co-pMG”, which stands for PSC derived microglia in co-culture with pNeuron, see Fig 1C rightmost panel and see the acronyms in Fig 1B). Haenseler teaches the embryonic-like macrophage precursors (i.e., PSC derived macrophage/microglia precursors) emerge into the supernatant as a uniform population of large, round cells with obvious filopodia and ruffles (p. 1728, right col, “Results” para 1). Thus, Haenseler, in view of Nestor, make obvious claim 6 (ii) continuing cultivation of the macrocarrier with the adherent embryoid bodies in cell culture medium until microglial precursor cells are released into the medium.
In regard to claim 6 (iii), Haenseler teaches the macrophage/microglia precursors can simply be harvested by collecting the supernatant without disrupting the EBs and replenishing flasks with fresh medium for many subsequent weekly harvests (p. 1728, right col, “Results” para 1), thus teaches claim 6 (iii) harvesting the microglial precursor cells released into the medium.
Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary.
Conclusion
No claims are allowed.
Examiner Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Jianjian Zhu whose telephone number is (571)272-0956. The examiner can normally be reached M - F 8:30AM - 4PM (EST).
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, James Douglas (Doug) Schultz can be reached on (571) 272-0763. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/JIANJIAN ZHU/Examiner, Art Unit 1631
/JAMES D SCHULTZ/Supervisory Patent Examiner, Art Unit 1631
Prosecution Insights