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 .
Claims Status
Claims 1 and 12 are amended.
Claim 8 is cancelled.
Claims 55 and 56 are newly added.
Claims 1-2,11-12,16,21-22, and 25-26 are under examination.
Withdrawn Objections
The objection raised against claim 12 is withdrawn in light of Applicants amendment.
Withdrawn Rejections
Claim Rejections - 35 USC § 102
The rejections of claims 1-2,8,11-12, 16, 21-22, and 25-26 under 35 U.S.C. 102(a)(1) as being anticipated by Bosch-Fortea et al (Biomaterials, 2019), hereafter Fortea et al. As evidenced by Azioune et al (Methods in Cell Biology, 2010) is withdrawn in light of Applicants amendment. Applicants amended claim 1 to recite an additional limitation reciting “ a leak-proof physical barrier attached to the first planar face, the leaky proof physical barrier physically separating respective sets of the plurality of microspots”. Applicants also amended claim 1 to recite “ wherein the three-dimensional aggregate of cells is an organoid”. The method of Fortea et al do not teach the aforementioned limitations. Therefore, the rejection is withdrawn.
Claim Rejections - 35 USC § 103
The rejections of claims 1-2, 8,11-12, 16,21-22, and 25-26 under 35 U.S.C. 103 as being unpatentable over Tewary et al (PLOS Biology, 2019) in view of Bosch-Fortea et al (Biomaterials, 2019), hereafter Fortea et al, is withdrawn in light of claim 1 amendment.
Response to Amendment
Applicant’s arguments have been carefully considered and found persuasive as
noted above. The new ground of rejection below addresses the deficiencies raised by
Applicant with respect to the amended claims.
New Grounds of Rejections
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 text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
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 1-2,11-12, 16,21-22, 25-26, and 55-56 are rejected under 35 U.S.C. 103 as being unpatentable over Tewary et al (PLOS Biology, 2019), in view of Wang et al (Biomaterials, 2009, Tsi et al ( BMC Cancer, 2018), and Bosch-Fortea et al (Biomaterials, 2019), hereafter Fortea et al.
Regarding claims 1, 12, and 55-56, Tewary et al disclose a method for culturing hPSCs to produce three-dimensional aggregates utilizing an anchorage dependent device. Tewary et al teach that the anchorage dependent device comprises a plurality of microspots created with PLL-g-PEG-based micropatterning on a glass substrate that is assembled to form a 96-well culture plate. Tewary et al used the photopatterning technology to create platforms with desired patterns of microspots ( i.e. circular, square, triangle, or rectangle). The method involves depositing the microspots to the surface of PEG-coated side of a coverslip by photo-oxidizing selected regions of the substrate using Deep UV exposure for 12 minutes through a Quartz photomask. According to Tewary et al, the disposal of microspots using Deep UV light and photomask (i.e. photopatterning technique) generates microspots with modified chemical attributes (i.e. carboxyl groups) that differ from the chemical attributes of the surrounding interstitial surface, with the microspots and not the interstitial surface supporting the tethering biological materials. Tewary et al also teach using carbodiimide and succinimide chemistry (EDC and NHS) to activate the carboxyl groups formed by the photomasking, allowing for covalent binding of ECM proteins, such as Geltrex basement membrane preparation, to the microspots. In one embodiment, Tewary et al demonstrate how to design microspots that are 200 microns in diameter with 500-microns separation. The separation distance reads on “ plurality of microspots separated by a pitch”. Tewary et al further disclose utilizing the device to assay hPSCs differentiation outcomes. ( See Fig.1-2, Materials and methods sections “ Preparation of PEG plates”, and “ Comparison between PEG plates with µCP plates”). In addition, the anchorage dependent device of Tewary et al comprises of micropatterned slide that are glued to a bottomless 96-well plates to produce microtiter plates with patterned cell-culture surfaces. It should be noted that the micropatterned slide reads on the first planar surface, whereas the bottomless 96-well plate reads on a leak-proof physical barrier attached to the first planar face. ( See Fig 1.B, and section “Preparation of PEG plates” on page 28). Tewary et al also describe utilizing the device to assay hPSCs differentiation outcomes. The method of Tewary involves culturing single cell suspension of hPSCs cells into the micropattern platform to produce three-dimensional cell aggregates, this reads on claim 56. ( See abstract, and Fig.1D). In particular, Tewary’s method for culturing PSCs involves suspending PSCs in SR medium supplemented with ROCK inhibitor and bFGF and then culturing the suspension onto the micropatterned platform for a period of 2 to 3 h till robust cell attachment is observed. When robust cell attachment is observed, ROCK inhibitor is removed from the media and then the cells are left overnight to make colonies. ( See section “ Comparison between PEG plates with μCP plates” on page 29). Tewary et al show that the micropatterned platform allows for the robust geometrical confinement of a variety of cell types in colonies of a variety of shapes and sizes. ( See Fig 1D).
Taken together, Tewary et al teach utilizing the micropatterned platform to grow PSCs into three-dimensional aggregates, but fail to teach inducing the PSCs into specific organoid.
Wang et al also teach utilizing a micropatterned-based platform to determine whether 3D multicellular spheroids of MSCs cultured in uniformly engineered microenvironments on the micropatterned platform would preserve multipotency and how it would impact the differentiation. Wang et al demonstrate that the micropatterned platform may be used to generate 3D MSCs spheroid of precise dimension and uniform quality, resulting in significantly improved cell differentiation. ( See abstract, Fig.2d, and section 3.1. on page 2707). According to Wang et al, “ Preciosity, reproducibility and homogeneity of this 3D spheroid culture platform proved to be excellent, a clear advantage in comparison to the conventional 2D monolayer and other 3D porous material counterparts”. Wang et al also state that “ This culture system is applicable for many scientific and medical fields that need outcomes in a high throughput manner”. ( See abstract, and 2nd column-2nd paragraph-on page 2707).
Tsi et al utilize the dome method to generate primary pancreatic tumor organoids from primary cells isolated from primary and metastatic human pancreatic tumors. The method involves dissociating the tumors into single cell suspension. The dissociated cells are then resuspended into liquefied Matrigel and plated in triplicate as droplets in the centers of wells of a 24-well tissue culture plate. Once the Matrigel droplets have solidified, the droplets are overlaid with organoid growth medium (OGM) with Rho kinase inhibitor to induce organoid formation. ( See section “ Organoid culture from primary tumor tissues and co-culture” on page 3-4, and Fig.1). Tsi et al teach that the generated primary pancreatic organoids are polarized with apicobasal polarity. ( See Fig.3c).
Taken together, claims 1,12, and 55-56 would have been obvious to one of ordinary skill in the art, as there was some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. Tewary et al utilize the micropatterned platform to grow hPSCs into three-dimensional cell aggregates, and clearly suggest that the platform enables robust geometrical confinement of a variety of cell types in colonies of varying shapes and sizes, but fail to suggest utilizing the platform to induce hPSCs to form a polarized organoids. Wang et al teach that the micropatterned platform can be utilized to generate 3D spheroids with precise dimensions and uniform quality, and clearly suggest that the platform can be utilized in many scientific and medical fields that need outcomes in a high throughput manner, but fails to suggest utilizing the platform to generate organoids. Tsi et al teach a method of differentiating primary pancreatic tumor cells into organoid using the dome method. Therefore, an ordinary skill in the art at the time the invention was filed who had reviewed Tewary et al could have come across Wang and Tsi and immediately noticed the benefit of modifying the teachings of Tewary et al to use the platform to produce polarized organoid with precise dimensions and uniform quality. On would be motivated to use the primary cells of Tsi et al or the MSCs of Wang et al and culture them on the platform of Tewary et al and then induce them to form organoids by using a supportive medium (i.e. OGM) as taught by Tsi et al to produce organoids with precise dimensions and uniform quality that can be used in a high throughput screening system such as to conduct drug discovery assays. See MPEP 2143 (I)(G).
Regarding claim2, the method of Tewary et al involves coating the micropatterned plates with Geltrex (diluted 1:150) for 4 h at room temperature on an orbital shaker. After incubation, the plate is washed with Phosphate Buffered Saline (PBS) at least 3 times to get rid of any passively adsorbed extracellular matrix (ECM).
Regarding claim 11, the combined teachings of Tewary, Wang, and Tsi render obvious claim 1. The instant claim recites a functional outcome “i.e. reduced off target cell differentiation between first and adjacent second three-dimensional aggregates grown using the anchorage surface”. This functional outcome is considered inherent, because the active step of the claim (i.e. culturing the cell suspensions on an anchorage dependent device comprising a plurality of microspots) is taught by Tewary et al in view of Wang et al and Tsi et al; and there is nothing in applicants' disclosure that show that these functional results comes from something other than the claimed method steps.
Regarding claim 16, Tewary et al teach utilizing the micropattern platform to expose the cell aggregates to different culturing conditions. For example, Tewary et al demonstrate that exposing undifferentiated hPSCs to different culturing conditions can drive their differentiation into a peri-gastrulation or preneurulation fate. Specifically, Tewary et al teach hPSCs can be induced into peri-gastrulation fate by culturing them in an SR medium supplemented with 100 ng/ml of bFGF and 50 ng/ml of BMP4 , and into preneurulation–like fate using an SR medium supplemented with 100 ng/ml of bFGF with 25 ng/ml of BMP4 and 10 μMSB431542. ( See Materials and methods section “Peri-gastrulation–like and preneurulation-like fate patterning induction”, 2nd paragraph, page 29).
Regarding claim 21, following the discussion above, the teachings of Tewary et al in view of Wang and Tsi render obvious claim 1. Tewary et al do not teach supplementing the culturing medium with sub-gelation dilution of the coating agent.
Fortea et al disclose a method of growing three-dimensional cellular structures composed of renal epithelial tubules using an anchorage dependent device comprising a plurality of microspots. (See Fig.1). Fortea et al teach that the generated tubules can be used in nephrotoxicity assay, and suggest that the anchorage dependent device is a powerful tool for studying the molecular mechanisms involved in organogenesis as well as conducting drug development assays. ( See Fortea abstract). The method of Fortea et al involves culturing cells in a supportive medium containing a sub-gelation concentration of Matrigel to produce polarized kidney-like tubules. According to Fortea et al, the growth of 3D structures relies on the coating patterns of the microspots with ECM proteins and supplementation of culture media with Matrigel. (See the 2nd column, 1st paragraph, on page 2). Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to modify the teachings of Tewary et al and use a supportive media containing a sub-gelation dilution of the coating agent, as taught by Fortea, to produce organoids. There would be a reasonable expectation of success because doing so would promote the formation of 3D organoids.
Regarding claim 22, Tewary et al also teach coating the microspots with soluble extracellular matrix proteins (ECM), such as Geltrex, which comprises of laminin, collagen IV, entactin, and heparin sulfate proteoglycan, as evidenced by Gibco data sheet. ( See Tewary et al Material and methods sections “ Preparation of PEG plates”, page 28) and (Gibco product sheet , 1st paragraph).
Regarding claim 25-26, Tewary et al demonstrate that photo-oxidizing organic polymers like PEG with Deep UV light for 12 minutes through a quartz photomask generates micropattern, carboxyl-rich regions. The presence of carboxylic group specifically on the microspots surface and not on the interstitial space generates microspots with a chemical attribute (i.e. carboxyl groups) that are more hydrophilic than the second chemical attribute of the interstitial space, which lacks such group due to the action of the photomasking. (See Fig.1).
Response to Arguments
Applicant's arguments filed 10/02/2025 have been fully considered but they are not
persuasive.
Applicants argue that neither Fortea nor Tewary teach utilizing the micropatterned platform to produce organoids from PSCs or primary cells. Applicants further argue that the anchorage dependent device of Fortea et al do not comprise a leak-proof physical barrier
attached thereto.
Examiner’s Response to Traversal: Applicant’s arguments have been carefully considered, but are not found persuasive. This is 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.
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 FATIMAH KHALAF MATALKAH whose telephone number is (703)756-5652. The examiner can normally be reached Monday-Friday,7:30 am-4:30 pm EST.
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/FATIMAH KHALAF MATALKAH/Examiner, Art Unit 1638
/Tracy Vivlemore/Supervisory Primary Examiner, Art Unit 1638