US-12619211-B2 - Method and apparatus for a tissue engineering system utilizing stacked layers

US12619211B2US 12619211 B2US12619211 B2US 12619211B2US-12619211-B2

Abstract

The present invention provides apparatus and methods for production of tissue structures and organs. In some examples, a cleanspace facility may be equipped with modelling hardware and software, nanotechnology and microelectronic apparatus, and additive manufacturing equipment to print cells and support matrix to allow cells to grow into tissue structures and organs. Various methods relating to using and producing the tissue engineering system are discussed.

Inventors

Frederick A. Flitsch
Robert A. Flitsch
Brent Chanin

Assignees

ORGANOFAB TECHNOLOGIES, INC.

Dates

Publication Date: 20260505
Application Date: 20210607

Claims (20)

1 . A method of forming a tissue layer comprising: configuring a tissue engineering apparatus comprising: a cleanspace fabricator, wherein the cleanspace fabricator is configured to process at least a first substrate, wherein the cleanspace fabricator maintains both a particulate cleanliness as well as a biological sterility cleanliness, wherein the cleanspace fabricator comprises at least a first processing apparatus and a second processing apparatus deployed along a periphery of the cleanspace fabricator, and wherein the cleanspace fabricator comprises automation to move one or more of the first substrate and the first processing apparatus within a primary cleanspace of the cleanspace fabricator; a modelling system, wherein the modelling system is configured to produce a first digital model which is used to control at least the first processing apparatus, wherein the first processing apparatus controls equipment to create one or more of a tissue support matrix and a printed deposit of cellular material and molecular material; and wherein the first processing apparatus comprises a support member with a multitude of printing elements arrayed thereupon, wherein the multitude of printing elements are capable of emitting a fluid comprising at least a first cell to a region within the first processing apparatus based upon a final three-dimensional model; and within the tissue engineering apparatus: placing a first deposit comprising a first cellular material upon the first substrate, wherein a location of the first deposit is controlled by the first digital model, and wherein the first substrate comprises a first material that is at least one of resorbable and dissolvable; placing a second deposit comprising a second cellular material upon a second substrate, wherein a location of the second deposit is controlled by the first digital model, and wherein the second substrate comprises a second material that is at least one of resorbable and dissolvable; positioning the second substrate upon the first substrate to form a stack, wherein a proximity of the second substrate to the first substrate allows the first deposit of the first cellular material to grow into a region of the second deposit of the second cellular material; and wherein the stack forms at least a portion of one of a tissue product or an organ product.
2 . The method of claim 1 further comprising: incubating the stack for an incubation period; and wherein during the incubation period, the first material that is at least one of resorbable and dissolvable is either resorbed within the cellular material or dissolved, respectively.
3 . The method of claim 1 wherein the first material is the same as the second material.
4 . The method of claim 1 wherein the first substrate comprises collagen.
5 . The method of claim 1 wherein the first processing apparatus further comprises a microfluidic processing system to process cellular and chemical material and deliver a product to the multitude of printing elements.
6 . The method of claim 1 wherein at least one of the multitude of printing elements is further used to print a third material comprising a chemical component, wherein the chemical component acts to signal cellular material.
7 . The method of claim 6 wherein the act to signal cellular material encourages the cellular material to adhere to the first substrate in the region where the third material is printed.
8 . The method of claim 6 wherein the act to signal cellular material encourages the cellular material to differentiate in a certain manner.
9 . The method of claim 1 wherein the placing of the first deposit includes a prior step wherein an alignment feature on the first substrate is detected by an element of the first processing apparatus to align the placing of the first deposit in space.
10 . The method of claim 9 wherein the positioning of the second substrate in a proximity to the first substrate includes a prior step, wherein an alignment feature on the first substrate is detected, and a prior step wherein an alignment feature on the second substrate is detected; and the detection of the alignment features are used to orient the positioning of the second substrate.
11 . The method of claim 1 wherein an algorithm to generate the first digital model uses medical imaging of a specific patient as an input.
12 . The method of claim 11 wherein the algorithm to generate the first digital model includes adjustments by a human operator to improve a function of the tissue layer or the organ product compared to an existing tissue or organ of the specific patient.
13 . The method of claim 11 wherein the algorithm utilizes an artificial intelligence algorithm to adjust the first digital model.
14 . The method of claim 13 wherein the artificial intelligence algorithm is trained on an atlas of imaging data of healthy individuals.
15 . The method of claim 14 wherein a generative design algorithm is used to adjust the first digital model to incorporate learned aspects of structure from the atlas of imaging data.
16 . The method of claim 1 wherein the first processing apparatus is used to print features after the stack is formed.
17 . A method of forming a tissue layer comprising: configuring a tissue engineering apparatus comprising: a cleanspace fabricator, wherein the cleanspace fabricator is configured to process at least a first substrate, wherein the cleanspace fabricator maintains both a particulate cleanliness as well as a biological sterility cleanliness, wherein the cleanspace fabricator comprises at least a first processing apparatus and a second processing apparatus deployed along a periphery of the cleanspace fabricator, and wherein the cleanspace fabricator comprises automation to move one or more of the first substrate and the first processing apparatus within a primary cleanspace of the cleanspace fabricator; and wherein the first processing apparatus comprises a support member with a multitude of printing elements arrayed thereupon, wherein the multitude of printing elements are capable of emitting a fluid comprising at least a first cell to a region within the first processing apparatus based upon a final three-dimensional model; within the tissue engineering apparatus: placing a first deposit comprising a first cellular material upon the first substrate, wherein a location of the first deposit is controlled by a first digital model; placing a second deposit comprising a second cellular material upon a second substrate, wherein a location of the second deposit is controlled by the first digital model; positioning the second substrate upon the first substrate to form a stack, wherein a proximity of the second substrate to the first substrate allows the first deposit of the first cellular material to grow into a region of the second deposit of the second cellular material; and wherein the stack forms at least a portion of one of a tissue product or an organ product.
18 . The method of claim 17 further comprising a modelling system, wherein the modelling system is configured to produce the first digital model which is used to control at least the first processing apparatus, wherein the first processing apparatus controls equipment to create one or more of a tissue support matrix and a printed deposit of cellular material and molecular material.
19 . The method of claim 17 further comprising incubating the stack for an incubation period.
20 . The method of claim 17 wherein the first substrate comprises collagen.

Description

CROSS REFERENCE TO RELATED APPLICATION This application claims priority to U.S. Non-Provisional Ser. No. 16/331,661 filed on Mar. 8, 2019 which in turn is the national stage filing of U.S. PCT Application S/N PCT/US2018/35984 which in turn claims the benefit of the U.S. Provisional Application Ser. No. 62/515,983 filed on Jun. 6, 2017. The contents of each are hereby incorporated by reference. FIELD OF THE INVENTION The present invention relates to methods and associated apparatus and methods which relate to fabrication systems, processing tools and modeling systems and protocols used to create tissue layers on substrates and organs. Arrays of multiple chemical species printing elements or cell printing elements may be combined with microfluidic processors and other techniques to form a tissue processing system. BACKGROUND OF THE INVENTION A cleanspace fabricator can create an environment that supports complex material processing in a simple clean environment that is also very sterile. In some examples, people are not located within the primary cleanspace of a cleanspace fabricator. Therefore, their cellular matter, and its associated DNA may be isolated as a contaminant for materials that are being processed in the cleanspace fabricator. There are many different processes that may be performed in a cleanspace fabricator which may benefit from the sterile and clean environment that it affords. Furthermore, there are numerous types of apparatus that may be created in a cleanspace environment such as the processing of microfluidic processing elements. Microfluidic processing elements may therefore be processed in a cleanspace fabricator and then be used in that cleanspace fabricator to perform processing themselves, leveraging the clean, genetically isolated, and sterile aspects of the environment. In nature, there are complex structures such as living tissues and organs that could be replicated or produced using technologies that could be efficiently operated within a cleanspace fabricator. The production of living tissues and organs could provide numerous benefits to medical needs of various kinds and to the field of regenerative medicine for example. A medical environment is an ideal place to study a patient with a medical imaging technique to determine shape, function, and abnormalities about various tissues and organ structures within a patient. The same environment is also an ideal place to extract tissue samples from a patient. A cleanspace facility could be figured to support operations within such a medical environment. In a clean and sterile environment, cells from tissue samples may be isolated and induced to grow into stockpiles of cells. Therefore, it would be very useful to create an environment that is sterile and well controlled, that may house and support equipment for the production of engineered tissues and organs. This may be especially useful if the cell stock that is used for the production of the engineered tissues and organs originates from a patient that requires the tissues or organs. Finally, it would also be useful if the information of medical imaging studies may be compiled to created models for the formation of the engineered tissues. SUMMARY OF THE INVENTION Accordingly, methods and apparatus for a tissue engineering system based on these principles are described herein. And the present invention provides apparatus and methods to create tissue layers on substrates and to create organs within a tissue engineering system that may be located within a cleanspace fabricator. Massively parallel implementations of chemical species printing elements or cell printing elements may be combined with other techniques to form a tissue processing system. The present invention may utilize modelling techniques to incorporate medical imaging data as well as functional knowledge of organs, tissue, and structure. In some examples, a model will calculate structures based on mathematical constructs, such as fractal equations, with constraints defined in other parts of the model and forms. The models may be used to form structure with techniques including photolithography, reactive ion etching, chemical etching, film deposition, additive manufacturing, and the like. The models may be used to detail locations to print different cell types, or different molecules which may attract different cell types to a location. In still further examples, the models may be used to print chemical and biochemical deposits that maybe used to direct differentiation of various pluripotent or omnipotent stems cells to form differentiated cell types. In some embodiments the tissue systems may be used in conjunction with cleanspace fabricators. Cleanspace fabrication facilities have been defined in various patent specifications, and the teachings and definition of these published specification may form a basis for understanding the utility of the inventive art herein within cleanspace environments. The present invention al