DETAILED ACTION
Claim Status
As of the Non-Final Office Action mailed 12/29/2025, claims 1-19 and 21 were pending and claim 21 was withdrawn for being drawn to nonelected invention.
In Applicant's Response filed on 3/30/2026, claims 1, and 4-5 were amended and claims 2-3 and 19 were canceled.
As such, claims 1, 4-18, and 21 are pending and claims 1 and 4-18 have been examined herein.
Withdrawn Objections/Rejections
The objections and rejections presented herein represent the full set of objections and rejections currently pending in this application. Any objections or rejections not specifically reiterated are hereby withdrawn.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 3/30/2026 is acknowledged. The IDS has been considered except where noted in the IDS. The Non-Patent Literature documents, Friedrich et al. and Stadler et al., were not considered because a copy was not provided to the Office.
Claim Interpretation
Claim 1 recites “wherein the cancer cells comprise non-spheroid-forming cells.” The examiner is interpreting this as conditional language because it is minimally limiting and the specification does not provide any characteristics for identifying this cell type, it does not describe what kinds of cells fall in this category. Moreover, depending on conditions the cell is put under, cells may or may not form spheroids. According to ThermoFisher Scientific (published 8/29/2024), cancer cells form spheroids when cultured in conditions that prevent cells from adhering to flat surfaces and by constraining their geometry, often using ultra-low adhesion U-bottom dishes. In some cases, spheroids can form without the addition of any extracellular matrix (ECM) proteins, while other models may require the addition of basement membrane extract (BME) or collagen to the media for more effective spheroid formation.
Claim 10-11 recite “inducing a vascular cell layer on the fibrotic layer” and “wherein the vascular cell layer is induced by endothelial or vascular endothelial cells”. The examiner is interpreting that the presence of endothelial or vascular endothelial cells is sufficient to induce a vascular cell layer.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claim(s) 1, 4, and 10-18 is/are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by Taniguchi et al (US20170285002A1, 3/16/2017; published 10/5/2017).
Taniguchi teaches method of forming cancer organoids (i.e., tumoroid) using primary cultured cancer cells derived from a cancer patient (abstract and para 3 of Taniguchi). The reference teaches coculturing a human pancreatic cancer cell line, human vascular endothelial cells (human umbilical vein endothelial cells: HUVECs), and human mesenchymal cells (human mesenchymal stem cells) (para 12 of Taniguchi) (“further comprising step (e) of inducing a vascular cell layer on the fibrotic layer” as in instant claim 10; “wherein the vascular cell layer is induced by endothelial cells or vascular endothelial cells” as in instant claim 11). The vascular endothelial cells can be collected from blood vessels or can be prepared according to a method known in the art from pluripotent stem cells such as induced pluripotent stem cells (iPS cells), embryonic stem cells (ES cells) or the like (para 59) (“wherein the endothelial cells or vascular endothelial cells are induced from induced pluripotent stem cells (iPSCs)” as in instant claim 12). The mesenchymal stem cells can be collected from bone marrow, fat tissue, placenta tissue, umbilical cord tissue, tissue of dental pulp, or the like, or can be prepared according to a method known in the art from pluripotent stem cells such as induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells) (para 60 of Taniguchi) (“A method for producing a fibrosis-encapsulated tumoroid, the method comprising steps of (a) obtaining cancer cells isolated from a subject, wherein the cancer cells comprise non- spheroid-forming cells; and (b) co-culturing the cancer cells with mesenchymal stem cells derived from induced pluripotent stem cells” as in instant claim 1; please note that, absent evidence to the contrary, the cancer cells of Taniguchi are non-spheroid forming cells (see claim interpretation above)). The ratio of co-cultured cells is not particularly limited so long as it permits cancer organoid formation but is preferably 10:1 to 100:1 (para 61) (“wherein the co-culture in step (b) is performed by mixing and culturing the cancer cells and the iPSC-derived MSCs at a cell number ration of 0.01:1 to 100:1” as in instant claim 4). The type of the cancer is not particularly limited and may be any cancer such as liver cancer, kidney cancer, malignant brain tumor, pancreatic cancer, stomach cancer, lung cancer or the like (para 58 of Taniguchi) (“wherein the cancer is a cancer selected from the group consisting of . . . stomach cancer, brain cancer, . . . , lung cancer, . . . kidney cancer, . . . , pancreatic cancer, . . . , liver cancer” as in instant claim 13). The medium for use in the culture may be any medium as long as the cancer organoid can be formed. For example, a medium for vascular endothelial cell culture, a medium for cancer cell culture, or a mixture of these two media is preferably used. Any medium for vascular endothelial cell culture may be used, and a medium containing at least one of hEGF (recombinant human epithelial cell growth factor), VEGF (vascular endothelial cell growth factor), hydrocortisone, bFGF, ascorbic acid, IGF1 (para 62). Any medium for cancer cell culture may be used, and examples thereof include DMEM medium (same para) (“wherein the co-culture is performed in at least one culture medium selected from the group consisting of DMEM” as in instant claim 14; “wherein the culture medium further contains . . . a growth factor” as in instant claim 16; “wherein the growth factor is at least one selected from the group consisting of VEGF, . . ., bFGF, EGF, and IGF-1” as in instant claim 17). The culture period is not particularly limited and is preferably 1 to 60 days, more preferably 1 to 7 days (para 69) (overlaps with “wherein the co-culture is performed for 2 days to 14 days” as in instant claim 18). Composition of culture solution described in working example 1 of Taniguchi contains B27 (para 103) (“wherein the culture medium further contains B27” as in instant claim 15).
Accordingly, absent evidence to the contrary, Taniguchi anticipates the invention of instant claims 1, 4, and 10-18.
Response to Arguments
Applicant’s arguments with respect to the previous rejection of claim(s) 1, 4-5, 13 and 18 have been considered but are moot because the new ground of rejection does not rely on any reference applied in the prior rejection of record for any teaching or matter specifically challenged in the argument.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
Claim(s) 1, 4-5, and 10-18 is/are rejected under 35 U.S.C. 103 as being unpatentable over Molla et al (J Tissue Eng Regen Med. 2018 Mar;12(3):727-737; Epub 4 Oct 2017) as evidenced by Harma et al (PLOS ONE 5(5): e10431; Published 3 May 2010) and in view of Taniguchi et al (US20170285002A1).
Molla teaches that human mesenchymal stem cells (MSCs) were seeded on the bone-mimetic scaffolds, where they differentiated into bone cells and then formed mineralized bone matrix without osteogenic supplements. Further, prostate cancer cells (cell line MDA PCa-2b) was seeded on MSCs-seeded scaffolds (see abstract and Materials and Methods). Sequentially cultured PCa cells with MSCs formed self-organized multicellular tumoroids with distinct tight cellular junctions and hypoxic core regions (Results para 1) (“A method for producing a fibrosis-encapsulated tumoroid, the method comprising steps of (a) obtaining cancer cells isolated from a subject, wherein the cancer cells comprise non- spheroid-forming cells; and(b) co-culturing the cancer cells with mesenchymal stem cells . . . to form a spheroid-shaped culture” as in instant claim 1 in-part; “wherein the cancer is a cancer selected from the group consisting of . . . prostate cancer” as in instant claim 13). For coculture studies, 5 × 104 hFOB cells and PCa cells were seeded simultaneously onto 3D scaffolds in 1:1 proportion. Similar coculture studies were performed for MSCs and PCa cells (“2.3 Cell Seeding”) (overlaps with “wherein the co-culture in step (b) is performed by mixing and culturing the cancer cells and the mesenchymal stem cells at a cell number ration of 0.01:1 to 100:1” as in instant claim 4).
Evidentiary reference Harma states prostate epithelial cells from both normal and cancer tissues, grown in three-dimensional (3D) culture as spheroids, represent promising in vitro models for the study of normal and cancer-relevant patterns of epithelial differentiation (abstract para 1). However, some cancer cell lines (VCaP, DuCaP, NCI-H660 and MDA-PCa 2b) failed to form spheroids, but persisted as single cells (“single” or “singular phenotype”) for up to 2 weeks. Thus, absent evidence to the contrary, the MDA-PCa-2b cell line of Molla is a non-spheroid-forming cancer cell.
The difference between Molla and the instant invention is that it does not teach that the mesenchymal stem cells are derived from induced pluripotent stem cells.
Taniguchi teaches method of forming cancer organoids (i.e., tumoroid) using primary cultured cancer cells derived from a cancer patient (abstract and para 3 of Taniguchi). The reference teaches coculturing a human pancreatic cancer cell line, human vascular endothelial cells (human umbilical vein endothelial cells: HUVECs), and human mesenchymal cells (human mesenchymal stem cells) (para 12 of Taniguchi) (“further comprising step (e) of inducing a vascular cell layer on the fibrotic layer” as in instant claim 10; “wherein the vascular cell layer is induced by endothelial cells or vascular endothelial cells” as in instant claim 11). The vascular endothelial cells can be collected from blood vessels or can be prepared according to a method known in the art from pluripotent stem cells such as induced pluripotent stem cells (iPS cells), embryonic stem cells (ES cells) or the like (para 59) (“wherein the endothelial cells or vascular endothelial cells are induced from induced pluripotent stem cells (iPSCs)” as in instant claim 12). The mesenchymal stem cells can be collected from bone marrow, fat tissue, placenta tissue, umbilical cord tissue, tissue of dental pulp, or the like, or can be prepared according to a method known in the art from pluripotent stem cells such as induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells) (para 60 of Taniguchi) (“co-culturing the cancer cells with mesenchymal stem cells derived from induced pluripotent stem cells” as in instant claim 1 in-part). The ratio of co-cultured cells is not particularly limited so long as it permits cancer organoid formation but is preferably 10:1 to 100:1 (para 61) (“wherein the co-culture in step (b) is performed by mixing and culturing the cancer cells and the iPSC-derived MSCs at a cell number ration of 0.01:1 to 100:1” as in instant claim 4). While Taniguchi does not explicitly describe a ratio of 1.5:1 to 2.5:1 as in instant claim 5, as per MPEP 2144.05(I), a prima facie case of obviousness exists where the claimed ranges or amounts do not overlap with the prior art but are merely close Titanium Metals Corp. of America v. Banner, 778 F.2d 775, 783, 227 USPQ 773, 779 (Fed. Cir. 1985). This is especially true here given that Taniguchi states that the ratio can be any ratio so long as cancer organoid formation occurs. Thus, it would have been a matter of routine experimentation using standard laboratory techniques available at the time of filing to determine the optimal co-culture cell number ratio as taught by Taniguchi with a reasonable expectation of success. The type of the cancer is not particularly limited and may be any cancer such as liver cancer, kidney cancer, malignant brain tumor, pancreatic cancer, stomach cancer, lung cancer or the like (para 58 of Taniguchi) (“wherein the cancer is a cancer selected from the group consisting of . . . stomach cancer, brain cancer, . . . , lung cancer, . . . kidney cancer, . . . , pancreatic cancer, . . . , liver cancer” as in instant claim 13). The medium for use in the culture may be any medium as long as the cancer organoid can be formed. For example, a medium for vascular endothelial cell culture, a medium for cancer cell culture, or a mixture of these two media is preferably used. Any medium for vascular endothelial cell culture may be used, and a medium containing at least one of hEGF (recombinant human epithelial cell growth factor), VEGF (vascular endothelial cell growth factor), hydrocortisone, bFGF, ascorbic acid, IGF1 (para 62). Any medium for cancer cell culture may be used, and examples thereof include DMEM medium (same para) (“wherein the co-culture is performed in at least one culture medium selected from the group consisting of DMEM” as in instant claim 14; “wherein the culture medium further contains . . . a growth factor” as in instant claim 16; “wherein the growth factor is at least one selected from the group consisting of VEGF, . . ., bFGF, EGF, and IGF-1” as in instant claim 17). The culture period is not particularly limited and is preferably 1 to 60 days, more preferably 1 to 7 days (para 69) (overlaps with “wherein the co-culture is performed for 2 days to 14 days” as in instant claim 18). Composition of culture solution described in working example 1 of Taniguchi contains B27 (para 103) (“wherein the culture medium further contains B27” as in instant claim 15).
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to produce a tumoroid using co-culture of MSCs and prostate cancer cells as taught by Molla, where the mesenchymal stem cells are derived from induced pluripotent stem cells as taught by Taniguchi, to arrive at the instantly claimed invention. Taniguchi shows that mesenchymal stem cells derived from induced pluripotent stem cells can successfully be used to produce cancer organoids, one of ordinary skill would have been motivated to simply substitute one known element (mesenchymal stem cells of Molla) for another (iPSC-derived MSCs of Taniguchi) to obtain the predictable result of advantageously producing a similar tumoroid capable of reproducing a cancer microenvironment with cancer stroma and reproduce cancer tissue similar to a structure in patients as taught by the prior art.
Claim(s) 6 and 8-9 is/are rejected under 35 U.S.C. 103 as being unpatentable over Taniguchi as applied to claims 1, 4, and 10-18 above, and further in view of Noel et al (J Vis Exp 2017 Aug 23; (126):56081; of record).
The difference between the teachings of Taniguchi and the invention as instantly claimed is that it does not teach inducing a fibrotic layer on the culture.
Noel teaches the preparation of 3-dimensional spheroid co-cultures of pancreatic cancer cells and fibroblasts (title). The reference teaches that many cancer types, including pancreatic cancer, have dense fibrotic stroma that plays a role in tumor progression and invasion (abstract). Activated cancer associated fibroblasts are a key component of the tumor stroma that interact with cancer cells and support their growth and survival (same para). Models that recapitulate the interaction of cancer cells and activated fibroblasts are important tools for studying the stromal biology and for development of antitumor agents (same para) (“inducing a fibrotic layer on the culture” as in instant claim 6; “wherein step (c) comprises encapsulating the fibrotic layer by at least one cell type selected from the group consisting of fibroblasts, . . . cancer-associated fibroblasts” as in instant claim 9). The reference teaches that once the spheroids are formed, they can be transferred for use for metabolic assays (“Washing and transfer of spheroids” para 1) (“further comprising step (d) of selecting a fibrosis encapsulated tumoroid on which the fibrotic layer has been induced” as in instant claim 8).
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to create a tumoroid as taught by Taniguchi, where the tumoroid has a fibrotic layer as taught by Noel, to arrive at the instantly claimed invention. As Noel shows that multi-cellular tumor spheroid containing fibroblasts can be used for metabolic assays and drug efficacy testing, one of ordinary skill would have been motivated to modify the method of Taniguchi to include a fibroblast layer as taught by Noel with a reasonable expectation of advantageously recapitulate the interaction of cancer cells and activated fibroblasts as taught by the prior art.
Response to Arguments
Applicant has not provided any arguments related to the Noel teachings as they relate to instant claims 6-8 nor has Applicant attempted to distinguish these claim from the teachings of Noel.
Claim(s) 7 is/are rejected under 35 U.S.C. 103 as being unpatentable over Taniguchi and Noel as applied to claims 6 and 8-9 above, and further in view of Leung et al (Biomater Sci. 2015 Feb; 3(2):336-44. Epub 2014 Nov 13; of record).
The teachings of Taniguchi and Noel in combination were recited in the above 35 U.S.C. 103 rejection as applied to claim 6 of which claim 7 depend. The teachings will not be repeated here.
The difference between the combined teachings and the invention as instantly claimed is that they do not teach compacting the induced fibrotic layer.
Leung teaches media additives to promote spheroid circularity and compactness (title). The reference teaches that comparing drug efficacy across different cell types in spheroid culture can be difficult due to variations in spheroid morphologies and transport characteristics (abstract). Improving the reproducibility of compact, circular spheroids contributes to standardizing and increasing the fidelity of the desired gradient profiles in these drug screening three-dimensional tissue cultures (same para). Multicellular spheroid models have been generated through various methods and has been applied to fabricate 3-D tissues such as cancer tumor models (Introduction para 2). Spheroids generally become more circular and compact when supplemented with low concentration of collagen compared to unsupplemented control conditions (“Effect of additives on spheroid morphology” para 3). Whereas high collagen concentrations had varied responses, primarily a negative impact on spheroid circularity and compactness and in some cell types (A549 and HeLa) completely abrogating single spheroid formation, similar to previously reported formation of small multi-spheroids within cultures containing higher concentrations of Matrigel (same para). In contrast, cells behave in a much more uniform fashion in the presence of MethoCel. For most cells tested (DU145, A549, HeLa, MDA-MB-231, MCF-7), the presence of MethoCel improved circularity and compactness of spheroids in a dose dependent manner with varying degrees of effectiveness (same para). MethoCel demonstrated its ability in enhancing spheroid morphology in the acute phase of spheroid culture (2 days), but it was unclear whether such enhancement would be sustained over a long culture period (“Effect of additives on spheroid morphology” para 4). To test this we chose three tumor cell lines (MDA-MB-231, DU145 and PC3) to further examine the effects of MethoCel on monoculture spheroid formation over a period of 4 days. Spheroids consisting of either MDA-MB-231 or DU145 became more circular and compact from day 1 to 4 post seeding (Fig. 3; same para). The addition of 0.24 mg mL−1 of MethoCel led to further enhancement over time when compared to cultures with no additives and spheroid morphology was minimally affected by culture time (same para).
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to create a tumoroid as taught by Taniguchi and Noel in combination, where the tumoroid is compacted as taught by Leung, to arrive at the instantly claimed invention. As Leung shows that spheroid morphologies and transport characteristics are effected by compactness and circularness, one of ordinary skill would have been motivated to combine the method of Taniguchi and Noel in combination with compacting via addition of medium supplements as taught by Leung according to known methods to yield the predictable result of advantageously improving the reproducibility of compact, circular spheroids and increase their fidelity as taught by the prior art.
Response to Arguments
Applicant has not provided any arguments related to the Leung teachings as they relate to instant claim 7 nor has Applicant attempted to distinguish this claim from the teachings of Ho.
Claim(s) 10-11 is/are rejected under 35 U.S.C. 103 as being unpatentable over Taniguchi and Noel as applied to claims 6 and 8-9 above, and further in view of Ho et al (Theranostics, 2012; 2(1):66-75; Epub 1 Jan 2012; of record).
The teachings of Taniguchi and Noel in combination were recited in the above 35 U.S.C. 103 rejection as applied to claim 6 of which claims 10-12 ultimately depend. The teachings will not be repeated here.
Ho teaches penetration of endothelial cell coated multicellular tumor spheroids (title). The reference teaches the investigation of the use of iron oxide nanoparticles that target tumor vasculature, via the tumstatin peptide, in a novel three-dimensional tissue culture model (abstract). The developed tissue culture model more closely mimics the in vivo environment with a leaky endothelium coating around a glioma tumor mass (same para). The novel endothelial cell-coated tumor model provides an in vitro microtissue environment to evaluate effects of drug therapy on neo-vascularization inhibition (same para). The reference teaches that Multicellular tumor spheroid (MTS), a three-dimensional cluster of cancer cells grown in vitro, mimics the early avascular stage of tumor growth such as the presence of ECM and diffusion gradients of nutrients and wastes (Introduction, para 2). As a result, MTS can contain heterogeneous regions of tumor cell growth including a necrotic center with proliferating cells restricted to the outer rim of the MTS (same para). A uniform in vitro tumor model comprising of both tumor spheroids and a vascular endothelium is highly desired to study the extravasation and penetration (Introduction para 4). They used micromolded non-adhesive agarose gels to construct a uniform MTS coated with endothelial cells (para 5). Utilizing the micromold and self-assembly techniques, cell-cell sorting interactions formed a model for a tumor core that must be accessed through a leaky vasculature (same para) (“further comprising step (e) of inducing a vascular cell layer on the fibrotic layer” as in instant claim 10; “wherein the vascular cell layer is induced by endothelial cells” as in instant claim 11). Rat RG2 cells, a glioblastoma model cell line, formed a MTS core similar to that of a solid tumor, while bovine-pulmonary arterial endothelial (BPAE) cells assembled on the surface of the MTS. The layer of endothelial cells restricted access to the tumor core and acted as the vascular endothelium (same para). This technique formed hundreds of uniform replicates for the endothelium-coated tumor (same para).
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to create a tumor spheroid as taught by Taniguchi and Noel in combination, where a vascular layer is formed on the surface of the spheroid as taught by Ho, to arrive at the instantly claimed invention. Ho shows endothelial cells can be utilized to create a vascular endothelial cell layer over a tumor spheroid. One of ordinary skill would have been motivated to modify the tumor spheroid of Taniguchi and Noel in combination to include an endothelial cell layer as taught by Ho with a reasonable expectation of advantageously creating a developed tissue culture model more closely mimics the in vivo environment with a leaky endothelium coating as taught by the prior art.
Response to Arguments
Applicant has not provided any arguments related to the Ho teachings as they relate to instant claims 10-11 nor has Applicant attempted to distinguish these claims from the teachings of Ho.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to GILLIAN C REGLAS whose telephone number is (571)270-0320. The examiner can normally be reached M-F 9-5.
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/G.R./Examiner, Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632
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