DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant's election without traverse of Group I, claims 1-13, and the species of a hydroxyl group (claim 1), general formula (2), R5 and R6 are hydrogen, R7 is an arbitrary organic group (claim 2), residue after a UV reaction (claim 3), claim 9 (claims 9, 10), in the reply filed on 26 March 2026 is acknowledged. Claims 10 and 14-21 have been withdrawn. Claims 1-9 and 11-13 are currently pending and under examination.
This Application is a national phase application under 35 U.S.C. §371 of International Application No. PCT/JP2022/018285, filed 20 April 2022, which claims priority to Japanese Patent Document No. JP2021-075266, filed 27 April 2021.
Claim Objections
Claim 1 is objected to because of the following informalities: “the hydrophilic polymer contain a phosphorylcholine group” should instead read “contains” (plural). It is noted that this grammatical error is also present in claims 2 and 3. Appropriate correction is required.
Claim 2 is objected to because of the following informalities: a closing “]” is present in the wherein clause, while the opening “[“ has been deleted; the closing bracket should be removed. 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 1-9 and 11-13 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 recites a “cell culture substrate comprising: a substrate”; this limitation is indefinite, because the term “substrate” is used to identify two different components. As such, it is unclear if the cell culture substrate comprises an additional substrate, or instead, if the second recitation of a substrate is intended to be the same substrate as the first recitation.
Further regarding claim 1, the limitation that the cell culture substrate “has regions (A) below and regions (B) below” is indefinite, because it is unclear what the term “below” is relative to. For example, below could mean underneath the substrate, below could mean in reference to each other (A and B), or below the layer.
Additionally regarding claim 1, the limitation of “an unevenness height at a boundary between each of the regions (A) and each of the regions (B) is 1 to 500 nm,” is indefinite, because it is unclear what “unevenness height” is intended to mean. For example, unevenness height indicates that a surface surrounding the boundary of each region is rough and varies by 1 to 500 nm, or the heights of regions (A) and regions (B) are intended to be different by 1 to 500 nm relative the whole of the region (e.g. the entirety of (A) is 1 nm higher than (B)). Further, it is unclear if only a boundary (i.e. the perimeter) of each region is intended to have an unevenness height, or instead if the whole of each region is intended to have the same height.
Claim 2 recites that “the hydrophilic polymer contain a compound represented by … General Formula (3) below”; this limitation is indefinite, because it is unclear if “represented by” indicates that the recited formula is merely exemplary, or instead, if “represented by” is intended to indicate that the compound must include the formula.
Claims 3-9 and 11-13 are included in this rejection, as these claims depend from above rejected claims and fails to remedy the noted deficiencies.
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.
Claims 1-9, 11, and 13 are rejected under 35 U.S.C. 103 as being unpatentable over Yamazaki et al. (A Novel Method to Prepare Size-Regulated Spheroids Composed of Human Dermal Fibroblasts, Biotechnology and Bioengineering, Vol. 44, (1994), pp. 38-44).
With regard to claim 1, Yamazaki et al. teach a cell culture system for preparing size-regulated spheroids (Abs.), which is a cell culture substrate, comprising a dish, which is a substrate, and a 2µm thick coating layer containing poly-N-isopropylacrylamide (PNIPAAm) conjugated with collagen (Abs.; p. 39, left col. to right col., Preparation of Coated dishes, 1-5), which is a layer containing a hydrophilic polymer with a layer thickness of 2000 nm covering at least a part of a surface of the substrate, the hydrophilic polymer containing a hydroxyl group. The coating layer on the dish has irradiated and nonirradiated areas (see Fig. 1), which are deemed to be regions (A) and regions (B) below.
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Referring to Fig. 5, reproduced here, each of the regions (A) are circular, which is island-shaped, and regions (A) have cell adhesion properties, while regions (B), which are the area surrounding the island-shaped regions (A), have no cell adhesion/cell proliferation properties (see Fig. 1).
The area of the island-shaped regions is 0.2 cm2 (20 mm2) for producing spheroids from human dermal fibroblasts (Abs.; Fig. 5). It is further taught that as the diameters of the photomask regions, which are the island-shaped regions, become smaller, the diameter of the spheroids become smaller, and viability increases (Fig. 4; Abs.). Consequently, Yamazaki et al. teach that size is important to cellular viability, with smaller diameter ranges taught to produce smaller spheroid size (350 µm) and increased cellular viability (Abs.). As such, it would have been obvious to one of ordinary skill in the art to decrease the area of the island-shaped regions to less than 20 mm2, as doing so would have been expected to provide spheroids with better viability. Further, it would have been routine for an ordinary artisan to determine the appropriate area of the adhesive islands based on the cell type being utilized and the desired spheroid size, which is regulated by the size of the regions.
While it is not specifically taught that there is an unevenness height at a boundary between each of the regions (A) and (B) of 1 to 500 nm, as the regions are intended to be soluble below a lower critical solution temperature, when that temperature is achieved during the dissolving process, there will be an unevenness in height throughout the regions as dissolution occurs.
It is noted that "the discovery of an optimum value of a variable in a known process is usually obvious." Pfizer v. Apotex, 480 F.3d at 1368. The rationale for determining the optimal parameters for prior art result effective variables "flows from the 'normal desire of scientists or artisans to improve upon what is already generally known.'" Id. (quoting In re Peterson, 315 F.3d 1325, 1330 (Fed. Cir. 2003)). Accordingly, it would have been obvious to optimize the area of the island-shaped regions, including to 0.001 to 5 mm2, and an unevenness height at a boundary between regions (A) and (B), including to 1 to 500 nm, to provide a cell culture system effective for the type of cell being utilized and the desired viability of the spheroid being produced, when using the taught system.
With regard to claim 2, Yamazaki et al. teach a cell culture system including the hydrophilic polymer poly-N-isopropylacrylamide (PNIPAAm) conjugated with collagen (Abs.; p. 39, left col. to right col., Preparation of Coated dishes, 1-5). Claim 2 is indefinite as discussed previously, and the noted formula is interpreted as merely exemplary. As such, Yamazaki et al. is deemed to teach a hydrophilic polymer as claimed.
With regard to claim 3, Yamazaki et al. teach that the hydrophilic polymer contains a monomer unit having a UV-reactive residue after a UV reaction (Fig. 1).
With regard to claim 4, Yamazaki et al. render obvious the cell culture substrate as claimed, including the components as claimed. As the cell culture substrate cannot be separated from its properties, the result that a ratio of a peak intensity at 287 eV to a peak intensity at 285 eV in a Cls spectrum of XPS measurement for the regions (A) is greater than a ratio of a peak intensity at 287 eV to a peak intensity at 285 eV in a Cls spectrum of XPS measurement for the regions (B) by 0.05 or more, would necessarily flow from the use of the components as rendered obvious.
With regard to claims 5 and 6, Yamazaki et al. teach a 2µm thick coating layer containing poly-N-isopropylacrylamide (PNIPAAm) conjugated with collagen (Abs.; p. 39, left col. to right col., Preparation of Coated dishes, 1-5; Fig. 1), which is a layer containing a temperature-responsive polymer, wherein the temperature-responsive polymer is a block copolymer containing a water-insoluble block segment and a temperature-responsive block segment. While it is not specifically taught that there is another layer containing a temperature-responsive polymer with a layer thickness of 1 to 100 nm on a surface of the layer containing the hydrophilic polymer, the duplication of a layer as already taught is obvious to one of ordinary skill in the art. Additionally, it is noted that "the discovery of an optimum value of a variable in a known process is usually obvious." Pfizer v. Apotex, 480 F.3d at 1368. The rationale for determining the optimal parameters for prior art result effective variables "flows from the 'normal desire of scientists or artisans to improve upon what is already generally known.'" Id. (quoting In re Peterson, 315 F.3d 1325, 1330 (Fed. Cir. 2003)). Accordingly, it would have been obvious to optimize the thickness of any layer utilized in the cell culture system as taught by Yamazaki et al., including to 1 to 100 nm, to provide a cell culture system effective for the type of cell being utilized and the desired viability of the spheroid being produced when using the system.
While it is not specifically taught that a ratio of a weight of the temperature-responsive block segment to a total weight of the water-insoluble block segment and the temperature-responsive block segment is greater than 90 weight%, it is noted that it is not inventive to find optimal workable ranges by routine experimentation. See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). As such, it would have been routine for an ordinary artisan to determine the appropriate ratio based upon the specific materials utilized and the cells to be cultured.
With regard to claim 7, Yamazaki et al. teach that the substrate is a plastic dish, specifically Falcon 3001 (p. 39, left col., Preparation of Coated dishes, 3.), which is made of polystyrene and has a base thickness of about 1.3mm (see Art of Record: University of Maryland). While it is not specifically taught that the substrate has a thickness of 0.01 to 0.5 mm, it would have been obvious to one of ordinary skill in the art to utilize a substrate having a desired thickness based upon the specific materials being utilized in the overall system and the specific cells to be cultured.
It is noted that Applicant indicates that polystyrene is a preferred material for the substrate (see Specification, para. 24). As such, the polystyrene substrate as taught by Yamazaki et al. has a refractive index of 1.4 to 1.6.
With regard to claim 8, as noted with regard to claim 7, Yamazaki et al. teach that the substrate is made of polystyrene and has a base thickness of about 1.3 mm, and Applicant indicates that polystyrene is a preferred material for the substrate (see Specification, para. 24). As such, the result that the fluorescence intensities of the substrate respectively excited at excitation wavelengths of 350 nm, 488 nm, and 647 nm are smaller than fluorescence intensities of a 1.2 mm thick polystyrene plate excited at the same excitation wavelengths, would necessarily flow from use of the system as taught by Yamazaki et al.
With regard to claim 9, Yamazaki et al. teach that the area of the island-shaped regions is 0.2 cm2 (20 mm2) for producing spheroids from human dermal fibroblasts (Abs.; Fig. 5). It is further taught that as the diameters of the photomask regions, which are the island-shaped regions, become smaller, the diameter of the spheroids become smaller, and viability increases (Fig. 4; Abs.). Consequently, Yamazaki et al. teach that size is important to cellular viability, with smaller diameter ranges taught to produce smaller spheroid size (350 µm) and increased cellular viability (Abs.). As such, it would have been obvious to one of ordinary skill in the art to decrease the area of the island-shaped regions to less than 20 mm2, as doing so would have been expected to provide spheroids with better viability. Further, it would have been routine for an ordinary artisan to determine the appropriate area of the adhesive islands based on the cell type being utilized and the desired spheroid size, which is regulated by the size of the regions.
In an embodiment, Yamazaki et al. teach 24 holes (regions (A)) on a 55 mm dish (Fig. 5). While it is not specifically taught that the number of regions (A) is 200 to 1,000 regions/cm2 based on a total area of the regions (A) and the regions (B), it is noted that it is not inventive to find optimal workable ranges by routine experimentation. See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). Here, it would have been routine for an ordinary artisan to determine the most appropriate number of adhesive regions on a single substrate based on the cells being cultured.
Additionally, it is noted that "the discovery of an optimum value of a variable in a known process is usually obvious." Pfizer v. Apotex, 480 F.3d at 1368. The rationale for determining the optimal parameters for prior art result effective variables "flows from the 'normal desire of scientists or artisans to improve upon what is already generally known.'" Id. (quoting In re Peterson, 315 F.3d 1325, 1330 (Fed. Cir. 2003)). Accordingly, it would have been obvious to optimize the area of the island-shaped regions, including to 0.005 to 0.2 mm2, and the number of regions based on a total area of (A) and (B), including to 200 to 1,000 regions/cm2, to provide a cell culture system effective for the type of cell being utilized and the desired viability of the spheroid being produced when using the taught system.
With regard to claim 11, Yamazaki et al. teach that a minimum distance between the regions (A) is about 3 mm (3,000 µM) (see Fig. 5C: scale bar is 5mm), which is fully encompassed within 500 to 10,000 µM.
With regard to claim 12, Yamazaki et al. render obvious the cell culture substrate as claimed, including the components as claimed. As the cell culture substrate cannot be separated from its properties, the system would necessarily be usable for inducing differentiation from pluripotent stem cells to three germ layer cells.
Claims 1 and 13 are rejected under 35 U.S.C. 103 as being unpatentable over Yamazaki et al., as applied to claim 1 above, and further in view of Caviedes et al. (US 2004/0219666; Published 2004).
The teachings of Yamazaki et al. as applied to claim 1 have been set forth above.
With regard to claim 13, while Yamazaki et al. teach a substrate made of polystyrene, it is not taught that the substrate is a polycarbonate or a cycloolefin polymer.
Caviedes et al. teach that cell culture substrates include plastics such as polystyrene or polycarbonate (Para. 14).
It would have been obvious to one of ordinary skill in the art to combine the teachings of Yamazaki et al. and Caviedes et al., because both teach cell culture substrate made of plastics, including polystyrene. The use of polycarbonate as a cell culture substrate is known in the art as taught by Caviedes et al. The use of polycarbonate in place of polystyrene as the substrate in the cell culture system of Yamazaki et al. amounts to the simple substitution of one known plastic cell culture substrate for another, and would have been expected to predictably and successfully provide an alternative plastic for use as the substrate in the system.
Conclusion
No claims are allowable.
Art of Record:
University of Maryland, Overview of culture dishes, Accessed: 9/15/2026, Available online at: biosciencecores.umd.edu/uploads/1/1/0/3/110331541/dishes.pdf (Falcon 3001 is made of polystyrene and has a base thickness of about 1.3mm).
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/JENNIFER M.H. TICHY/Primary Examiner, Art Unit 1653
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