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
Applicant’s election without traverse of Inventive Group I in the reply filed on 07/07/2025 is acknowledged.
Claims 9 and 16-24 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 07/07/2025.
Applicant is reminded that upon the cancelation of claims to a non-elected invention, the inventorship must be corrected in compliance with 37 CFR 1.48(a) if one or more of the currently named inventors is no longer an inventor of at least one claim remaining in the application. A request to correct inventorship under 37 CFR 1.48(a) must be accompanied by an application data sheet in accordance with 37 CFR 1.76 that identifies each inventor by his or her legal name and by the processing fee required under 37 CFR 1.17(i).
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.
Claims 1, 3, and 15 are rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006) as evidenced by the Merriam-Webster definition of “aggregate” and “room temperature”
Regarding Claim 1: Gage teaches a method of culture that results in the generation of neuroblasts (0015-0006) which form aggregates comprising a “extensive network of processes forming a lattice-type pattern” in the presence of bFGF. (0071) Gage further teaches that the neuroblasts of the claimed invention can be stored in the vapor phase of liquid nitrogen when a cryopreservation agent is present. (0023) This reads on the claimed method of freezing cell aggregates comprising neural cells with a three-dimensional structure. Gage fails to teach soaking the aggregates in a cryopreservation solution prior to freezing and the claimed temperature range of the nitrogen storage unit.
Ghetler teaches a method of slow-freezing cryopreservation for early embryos and zygotes that involves incubation of the zygote/embryo for 10 minutes at 37°C followed by two soaking steps carried out at room temperature for 10 minutes each in the cryopreservation medium prior to a series of cooling down steps ultimately resulting in storage in liquid nitrogen at a temperature of -196°C. (Pg 211, Cryopreservation Protocol) This reads on the claimed method of soaking the cell aggregate at a temperature between 0°C-320°C prior to cryopreservation at a temperature of -150°C or less, as the Merriam-Webster medical definition of room temperature is a temperature range of 15-25°C.
Ghetler teaches a cryopreservation method of sequential soaking steps for the aggregates in the cryopreservation solution which comprises three sequential incubation steps at room temperature for 10 minutes each. As the incubation steps at room temperature comprise 30 minutes total, this reads on the claimed method of soaking the cell aggregate in the cryopreservation solution for 15-360 minutes.
Regarding Claim 3: Gage teaches that storage for long periods of time can be achieved in the vapor phase of liquid nitrogen. (0023) Gage fails to specify that the cryopreservation solution-soaked cell aggregate contained in the liquid nitrogen container has a volume that is 5% or less than the volume of the vapor phase of the liquid nitrogen container.
Ghetler teaches that the cell aggregates of the claimed invention are cryopreserved in open pooled straws, which are known in the art to have a volume of less than 1mL and are commonly used in oocyte cryopreservation. (Pg 1, introduction) As this is such a small volume of liquid comprising the cell aggregate, it would be obvious to a person skilled in the art that the volume of the aggregate contained within the open pool straw taught by Ghetler would be under 5% of the total volume of the vapor phase nitrogen storage unit.
Regarding Claim 15: Gage teaches that nitrogen vapor is used for the long-term storage of cell lines when a cryopreservation agent is present. (0023) Furthermore, as discussed above, Ghetler teaches soaking of the cell aggregate for a total of 30 minutes in the cryoprotectant solution at room temperature, reading on soaking at a temperature range of 0-20°C and Gage teaches storage of neuroblasts (a cell aggregate) in the vapor phase of a liquid nitrogen storage unit.
In addition to this, as discussed above the teachings of Ghetler read on the claimed range of 0-20°C as evidenced by the Merriam-Webster dictionary definition of “room temperature”. The storage of neuroblasts as taught by Gage reads on the claimed method of the cell aggregate having a three-dimensional structure (0086) which when combined with the teachings of Gage of use of nitrogen vapor (0023) result in a storage system in which soaks the cell aggregates in a cryopreservation solution at a temperature range of 0-20°C for 15-360 minutes prior to freezing which is then cryopreserved in the vapor phase of liquid nitrogen having a temperature of -150°C or less.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the neuroblast aggregate culture method of Gage with the cryopreservation protocol taught by Ghetler to create a method for freezing a neural aggregate comprising soaking the cell aggregate in a cryopreservation solution prior to freezing in the vapor phase of liquid nitrogen. One would have been motivated and had a reasonable expectation of success due to the sequential incubation steps of soaking the aggregates in cryopreservation media at room temperature as taught by Ghetler and the teachings of Gage that state nitrogen vapor storage is useful for long-term cell cryopreservation as long as there is a cryopreservation agent present.
Claims 4 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006) and StemCell Technologies (Intestinal Epithelial Organoid Culture with IntestiCult™ Organoid Growth Medium (Mouse), 2016)
The teachings of Gage and Ghetler are disclosed above. Gage and Ghetler fail to teach the claimed filling density of 50-500 cell aggregates per mL of cryopreservation solution or that the total volume of the cell aggregates and cryopreservation solution is between 0.5-5mL.
Regarding Claim 4: StemCell Technologies teaches that the recommended number of organoids for cryopreservation is 200 organoids per cryovial. (Pg 7, Section 4: Cryopreservation of Mouse Intestinal Organoids) This reads on the claimed range of 50-600 cell aggregates/mL.
Regarding Claim 5: StemCell Technologies teaches that the organoid pellet, after centrifuging, should be resuspended in 1mL of cryopreservation media per 1 cryovial of 200 organoids. (Pg 7, Section 4: Cryopreservation of Mouse Intestinal Organoids) This reads on the claimed range of cryopreservation solution being 0.5-5mL.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the teachings of StemCell Technologies regarding cryopreserving 200 organoids in 1mL of cryopreservation solution with the neural organoid culture method taught by Gage and the slow cryopreservation method taught by Ghetler to cryopreserve 200 organoids in 1mL of cryopreservation media. One would have been motivated and had a reasonable expectation of success at doing so based on the teachings of StemCell Technologies of optimal results being obtained at cryopreservation with 200 organoids per 1mL of cryopreservation media.
Claims 6-8 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006) and Pan et al. (US 2019/0046583 A1)
The teachings of Gage and Ghetler are disclosed above. Gage and Ghetler fail to teach use of stem-cell derived neural cells which express FOXA2 or TH or NURR1, and LMX1A.
Regarding Claim 6: Pan teaches that the brain microphysiological system aggregate (BMPS) can be produced from induced pluripotent stem cells. (0004)
Regarding Claims 7 and 8: Pan teaches an embodiment of the invention in which the neural cell types of the aggregates express FOXA2, TH, and LMX1A. (0009)
Regarding Claim 12: Pan teaches an embodiment of the invention in which the spheroid mass has a diameter of less than 500µM in diameter. (0010) This reads on the claimed range of the neural cell aggregate having a diameter between 150µM and 1000µM.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the specifications taught by Pan regarding the size of the organoid, the markers it expresses, and the source the cells are from with the neural organoid culture method taught by Gage and the slow cryopreservation method taught by Ghetler to create a neural organoid derived from stem cells which expresses FOXA2, TH, and LMX1A and is between 150µM and 1000µM in size. One would have been motivated and had a reasonable expectation of success at doing so due to the teachings of Pan, who state that neural organoids can be less than 500µM in diameter, derived from iPSCs, and that they can express FOXA2, TH, and LMX1A, which are all neural biomarkers.
Claims 10 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006) and Smits et al. (Modeling Parkinson’s disease in midbrain-like organoids, 2018)
The teachings of Gage and Ghetler are disclosed above. Gage and Ghetler fail to teach that at least 60% of the cells of the aggregates are dopamine-producing neuron progenitor cells.
Regarding Claim 10: Smits teaches a method for generation of organoids which resemble the complexity of the human midbrain comprised of dopamine producing midbrain dopaminergic neurons (mDANs). (Pg 1, Abstract)
Regarding Claim 11: Smits teaches that high-content image analysis at day 35 of culture showed that 62.38% +/- 3.5% of the cells in the human midbrain-specific organoids were differentiated into mDANs. (Pg 2, Results) This reads on the claimed method of at least 60% or more of the cell aggregate being comprised of dopamine-producing neuron progenitor cells.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to use the differentiation method taught by Smits which resulted in dopamine producing neural progenitor cells with the organoid culture method taught by Gage and the slow cryopreservation method taught by Ghetler to create a cell aggregate for cryopreservation that consists of 60% or more dopamine-producing neural progenitor cells. One would have had motivation and a reasonable expectation of success due to the teachings of Smits, who demonstrate that over 60% of the cells in their generated organoids were dopamine producing midbrain dopaminergic neurons
Claim 13 is rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006) and Xie et al. (Organoid Culture of Isolated Cells from Patient-derived Tissues with Colorectal Cancer, 2016)
The teachings of Gage and Ghetler are disclosed above. Gage and Ghetler fail to teach that the cell aggregate includes 500-150000 cells.
Regarding Claim 13: Xie teaches a method of generation of tumor organoids and compares different seeding densities for the generation of said organoids. This was due to the fact that organoids have a necrotic core, and an optimal size is necessary for the establishment of physiological gradients for pH, oxygen, nutrients, and waste to ensure optimal health of the cultures. Xie teaches that the organoids generated from 2000 cells performed the best, however all seeding densities formed organoids. (Pg 2471, Results) This reads on the claimed range of use of 500-150000 cells to form the cell aggregates.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to use the teachings of Xie of 2000 cells per organoid with the organoid culture method taught by Gage and the slow cryopreservation method taught by Ghetler to cryopreserve a cell aggregate comprising 500-150000 cells per aggregate. One would have been motivated and had a reasonable expectation of success at doing so based on the teachings of Xie, who state that organoids seeded from 2000 cells/well provide an adequate cultivate system and grew 8-fold over 25 days. (Pg 2471, Results)
Claim 14 is rejected under 35 U.S.C. 103 as being unpatentable over Gage et al. (US 2002/0039789 A1) in view of Ghetler et al. (Human oocyte cryopreservation and the fate of cortical granules, 2006), Xie et al. (Organoid Culture of Isolated Cells from Patient-derived Tissues with Colorectal Cancer, 2016), and StemCell Technologies (Intestinal Epithelial Organoid Culture with IntestiCult™ Organoid Growth Medium (Mouse), 2016)
The teachings of Gage and Ghetler are disclosed above. Gage and Ghetler fail to teach that the number of cells contained in the cryopreservation solution is 80000-5000000 cells/mL.
Regarding Claim 14: StemCell Technologies teaches the recommended number of organoids for cryopreservation is 200 organoids per cryovial (pg 7, Section 4: Cryopreservation of Mouse Intestinal Organoids) and that the organoid pellet, after centrifuging, should be resuspended in 1mL of cryopreservation media per 1 cryovial of 200 organoids. (Pg 7, Section 4: Cryopreservation of Mouse Intestinal Organoids) StemCell Technologies fails to teach use of 80000-5000000 cells/mL for cryopreservation.
Xie teaches that the organoids generated from 2000 cells performed the best and that the organoids seeded from 2000 cells/well provide an adequate cultivate system and grew 8-fold over 25 days. (Pg 2471, Results) Use of 2000 cells per organoid as taught by Xie combined with the teaching of use of 200 organoids per 1mL of cryopreservation solution per vial as taught by StemCell Technologies results in a total cell density per mL of 400000 cells/mL contained in the cryopreservation solution, which reads on the claimed range of 80000-5000000 cells/mL.
It would have been obvious to a person of ordinary skill in the art before the effective filing date of the claimed invention to combine the teachings of StemCell Technologies of 200 organoids per 1mL of cryopreservation solution per cryovial and the teachings of Xie of 2000 cells/well for the formation of a robust organoid with the organoid culture method taught by Gage and the slow cryopreservation method taught by Ghetler to create a cell aggregate cryopreservation method using 80000-5000000 cells/mL of cryopreservation media. One would have been motivated and had a reasonable expectation of success at doing so due to the recommendation of 200 organoids per mL of cryopreservation solution taught by StemCell Technologies and the recommended cell seeding density of 2000 cells per well for the generation of organoids as taught by Xie.
Response to Arguments
Applicant's arguments filed 10/21/2025 have been fully considered but they are not persuasive. The Applicant’s primary argument is that the soaking step taught by Ghetler is carried out at room temperature, and the amended range for soaking prior to cryopreservation is that of 0-20°C. However, this is unpersuasive because the established definition of “room temperature” as per Merriam-Webster is that of a temperature between 15°-25°C, or one that is suitable for human occupancy. (Merriam-Webster, Medical Definition of “Room Temperature”) As the accepted range for room temperature used by Ghetler of 15°-25°C overlaps the amended claimed range of 0-20°C, a prima facie case of obviousness exists as stated in the MPEP (2144.05.I):
“In the case where the claimed ranges "overlap or lie inside ranges disclosed by the prior art" a prima facie case of obviousness exists. In re Wertheim, 541 F.2d 257, 191 USPQ 90 (CCPA 1976); In re Woodruff, 919 F.2d 1575, 16 USPQ2d 1934 (Fed. Cir. 1990) (The prior art taught carbon monoxide concentrations of "about 1-5%" while the claim was limited to "more than 5%." The court held that "about 1-5%" allowed for concentrations slightly above 5% thus the ranges overlapped.); In re Geisler, 116 F.3d 1465, 1469-71, 43 USPQ2d 1362, 1365-66 (Fed. Cir. 1997) (Claim reciting thickness of a protective layer as falling within a range of "50 to 100 Angstroms" considered prima facie obvious in view of prior art reference teaching that "for suitable protection, the thickness of the protective layer should be not less than about 10 nm [i.e., 100 Angstroms]." The court stated that "by stating that ‘suitable protection’ is provided if the protective layer is ‘about’ 100 Angstroms thick, [the prior art reference] directly teaches the use of a thickness within [applicant’s] claimed range."). See also In re Bergen, 120 F.2d 329, 332, 49 USPQ 749, 751-52 (CCPA 1941) (The court found that the overlapping endpoint of the prior art and claimed range was sufficient to support an obviousness rejection, particularly when there was no showing of criticality of the claimed range).”
Therefore, the teachings of Ghetler still read on the newly claimed range of 0-20°C and the rejection on claim 1 is upheld. Furthermore, the incubation steps as taught by Ghetler are for 10 minutes each and comprise two 10 minute incubations at room temperature, reading on the 15-360 minute range for the 0-20°C incubation step as claimed. Therefore, the argument presented by the Applicant that the combined teachings of Gage and Ghetler fail to read on the claimed time range of 15 to 360 minutes for the incubation step is unpersuasive. In addition to this, Applicant has argued that Gage fails to teach the generation of aggregates and that Ghetler, who teaches use of zygotes and early embryos, fails to remedy the deficiency in teachings. However, the definition of the word per Merriam-Webster of “aggregate” is “a group, body, or mass composed of many distinct parts”. Therefore, a blastocyst comprising of a collection/aggregation of cells does read on the claimed “cell aggregate”.
Based on the above discussion, the rejection of claim 1 is upheld.
The arguments regarding claims 3-8 and 10-15 all rely on the argument set forth on claim 1, which per the above discussion failed to be persuasive. Therefore, the rejections for claims 3-8 and 10-15 are upheld.
Conclusion
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 extension fee 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 HANNA M THUESON whose telephone number is (571) 272-3680. The examiner can normally be reached M-F 7:30-5 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, Tracy Vivlemore, can be reached on (571) 272-2914. 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.
/HANNA MARIE THUESON/ Examiner, Art Unit 1638
/Tracy Vivlemore/Supervisory Primary Examiner, Art Unit 1638