This disclosure relates generally to an apparatus and method for processing tissue. More specifically, this disclosure relates to isolating tissue, for example, from tissue samples or organs by way of a tissue processing chamber.
Many different methods and approaches have been attempted to isolate individual cells from their respective parent organs or larger tissue samples. Prior methods have produced isolated cells with some cell destruction. This cell destruction can result from the relatively severe mechanical stimulation that is used to isolate cells from an organ. Additionally, many known methods require addition of an enzyme to break down the tissue samples.
The disadvantages of mechanical and enzymatic methods for individual cell isolation from parent organs or tissues known in the art has resulted in a need in the art for more effective devices and methods for individual cell isolation from parent organs or tissues that provides greater yields of a greater percentage of intact, viable cells.
Provided herein are devices and methods of use thereof for individual cell isolation from parent organs or tissues that provides greater yields of a greater percentage of intact, viable cells.
One aspect of the disclosure is a tissue processing device. The tissue processing device includes a tissue chamber. The tissue chamber includes at least one rotary blade housed within the tissue chamber, a drive shaft coupled to the at least one rotary blade, wherein rotation of the drive shaft is configured to rotate the at least one rotary blade, and a screen adjacent to the rotary blades, wherein rotation of the at least one rotary blade is configured to press processed tissue of a tissue sample through the screen. The tissue processing device further includes a collection chamber coupled to the tissue chamber configured to collect the processed tissue.
In another aspect, a tissue processing system is disclosed. The tissue processing system includes a tissue chamber. The tissue processing chamber includes at least one rotary blade housed within the tissue chamber, a drive shaft coupled to the at least one rotary blade, wherein rotation of the drive shaft is configured to rotate the at least one rotary blade, and wherein a distal end of the drive shaft comprises a motor coupling, and a screen adjacent to the at least one rotary blade, wherein rotation of the at least one rotary blade is configured to press processed tissue of a tissue sample through the screen. The tissue processing system further includes a collection chamber coupled to the tissue chamber configured to collect the processed tissue and an isolation chamber coupled to the tissue chamber and the collection chamber. The isolation chamber includes a motor coupled to the motor coupling configured to rotate the drive shaft
In another aspect, a tissue processing method is disclosed. The tissue processing method includes rotating at least one rotary blade within a tissue chamber. The tissue processing method additionally includes pressing at least a portion of a tissue sample through a screen adjacent to the at least one rotary blade via impeller forces of the at least one rotary blade. The tissue processing method further includes collecting processed tissue in a collection chamber.
These and other features and advantages of the invention disclosed herein will be more fully understood from the following detailed description taken together with the accompanying drawings and the claims. It is noted that the scope of the claims is defined by the recitations therein and not by the specific discussion of features and advantages set forth in the present description.
The above, as well as additional, features will be better understood through the following illustrative and non-limiting detailed description of example embodiments, with reference to the appended drawings.
All the figures are schematic, not necessarily to scale, and generally only show parts which are necessary to elucidate example embodiments, wherein other parts can be omitted or merely suggested.
Example embodiments are now described more fully hereinafter with reference to the accompanying drawings. That which is encompassed by the claims can, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided by way of example. Furthermore, like numbers refer to the same or similar elements or components throughout.
In accordance with the principles herein, a tissue processing chamber, shown generally at 100, provides processing and separation of tissue samples from larger tissue samples or organs. The tissue processing chamber can include rotary blades which, through impeller forces of the rotating rotary blades, press the larger tissue sample through a screen into a collection chamber. Processed tissue can then be extracted from the collection chamber, for example, for testing, culture, or clinical use.
For example, tissues that can be processed include, but are not limited to mesodermal-derived, endodermal-derived, ectodermal derived tissues, extraembryonic and fetal adnexa tissues, adipose tissue, pancreatic tissue, liver tissue, biliary tree issue, intestinal tissue, lung tissue, kidney tissue, bone tissue, bone marrow tissue, cartilage, muscle tissue, tendon, ligaments, amniotic tissue, chorionic tissue, umbilical cord tissue, placenta, blood vessels tissue, ovarian tissue, endocrine tissue, thyroid gland tissue, parathyroid gland tissue, adrenal gland tissue, pituitary gland tissue, pineal gland tissue, thymic tissue, dermal tissue, epidermal tissue, connective tissue, fibrous tissue, and central and peripheral nervous tissue. Tissue processing can be performed by activating the impeller at a specific rotational speed, or at a range of speeds, in clockwise or counter-clockwise direction. The geometry of the blade and of the screen can be modified to yield tissue fragments with different shapes. Screens of different geometries can be replaced in the same instrument to yield tissue fragments of different sizes. Instruments connected in series and loaded with screens of progressively smaller sizes can process tissue yielding fragments of decreasing size throughout the series.
Further, testing can include, but is not limited to, measurement of the size, volume, and number of tissue fragments via imaging, measurement of the weight of the fragments via mechanical scale or balance, measurement of electrical impedance of tissue fragments, suspended in an electrolyte, when passing through an aperture between electrodes (Coulter method, Coulter principle), measurement of viability of tissue fragments via staining and imaging, analysis of RNA expression and gene expression via northern blotting, hybridization, fluorescent in situ hybridization, reverse transcription-Polymerase Chain Reaction (RT-PCR), quantitative RT-PCR, microarray, Tiling array, next-generation sequencing, RNA sequencing, analysis of DNA content via DNA sequencing, analysis of protein expression via Liquid Chromatography—Tandem Mass Spectrometry, Gas Chromatography, analysis of immunomodulatory function, analysis of hormone-release function, and/or analysis of the release of factors in the fluid milieu via sensors.
In some example embodiments, culture that can be done with samples include tissue fragments can be cultured with culture media to generate organotypic cultures; tissue fragments can be cultured alone or in combination with other cells, tissue fragments, tissues, or matrices; tissue fragments can be cultured with culture media in static culture, in agitation, in perifusion (i.e., fluid flow), in an automated bioreactor system; and/or in two-dimensional or three-dimensional culture conditions, or in a compartimentalized device. Tissue fragments can be cultured in culture media in non-adherent conditions, in adherent conditions, or in embedded conditions (such as in a matrix or material); tissue fragments can be maintained in a liquid medium, or at the liquid-gas interface; tissue fragments can be suspended in cryopreservation medium and subsequently cryopreserved, can be cryopreserved directly, can be lyophilized, can be maintained in hypothermic conditions, or can be encapsulated.
In some example embodiments, clinical uses for the samples include, but are not limited to, manipulation of the tissue via mechanical processing of tissue into tissue fragments, in the presence or absence of washing, concentration, and/or preservation steps. Minimally manipulated tissue can be utilized for homologous use and can be utilized clinically in the autologous setting, or in the allogeneic setting. Processed tissue fragments can be implanted for homologous use (i.e., for repair, reconstruction, replacement, or supplementation of a recipient's cells or tissues). In homologous uses, fragments of tissue can perform the same basic function or functions in the recipient as in the donor. Human tissues undergoing minimal manipulation and intended for application in homologous uses are currently classified as human cellular or tissue product (HCT/P). Adipose tissue fragments can be utilized in plastic surgery, musculoskeletal, reparative (traumatic lesions, burns and wounds) regenerative medicine applications, and reconstructive surgery applications. Cartilage tissue fragments can be utilized in reconstructive and orthopedic surgery application to replace cartilage after fracture, loss, or disease. Stromal Vascular Tissue Fragments can be utilized in reconstructive surgery applications. Endocrine tissue fragments can be used to functionally replace or supplement the endocrine tissue of the recipient. Optionally, processed tissue fragments can be cultured and/or cryopreserved, before clinical use.
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In example embodiments, the tissue chamber 102 can be cylindrical, or substantially cylindrical, and house a portion of the drive shaft 104, the rotary blades 106, the support grid 108, and the screen 112. More specifically, the tissue chamber 102 can be a vessel defined by an outer boundary and a space within the outer boundary. The space within the outer boundary can have any useful and convenient shape. Example configurations include cylindrical (as shown in
In some examples, the tissue chamber 102 can be constructed of an autoclavable material. Autoclavable material can withstand the pressure and temperature of tissue processing, as well as repeated sterilization. For example, the tissue chamber 102 can comprise a high grade polymer material. This is desirable, as tissue processing requires regulated temperatures and pressures. It should be understood that other materials and example configurations of the tissue chamber 102 are possible.
The tissue chamber 102 includes an inlet for depositing a tissue sample, such as a tissue loading port 123. The tissue loading port 123 can include a tissue loading port cap 125. The tissue loading port 123 can be configured such that, during use, the tissue chamber 102 can be assembled and sterilized before a tissue sample is added. The tissue sample can then be added by removing the tissue loading port cap 125 and depositing the tissue sample into the tissue chamber 102. In some example, the tissue loading port 123 and tissue loading port cap 123 can fasten to each other by way of a threaded connection, however other example embodiments are possible.
Similar to the tissue chamber 102, in some examples, the tissue loading port 123 and tissue loading port cap 125 can include autoclavable material, such as a high grade polymer material. Additionally or alternatively, the tissue loading port 123 and tissue loading port cap 125 can include material that can be sterilized via irradiation or via gas sterilization. It should be understood that other materials and example configurations of the tissue loading port 123 and tissue loading port cap 125 are possible.
Additionally or alternatively, the tissue sample can be deposited into the tissue chamber 102 directly, for example, from a top portion of the tissue chamber. In an alternative embodiment, the tissue sample can be deposited into the tissue chamber 102 before the tissue chamber 102 is coupled to the collection chamber 110. For example, the tissue chamber 102 and collection chamber 110 can include a threaded connection 127 for deposit and removal of the tissue sample, as shown in
Additionally or alternatively, the tissue sample can be deposited by way of an inlet, such as a luer lock 114, or equivalent. The luer lock 114 can include fluid fittings used for making leak-free, sterile connections between a male-taper fitting and its mating female part on the tissue chamber 102. The luer lock 114 can couple to an inlet tube (not shown) to deposit a specimen, such as homogenate, or saline into the tissue chamber 102. Additionally, or alternatively, in some examples, the inlet tube coupled to the luer lock 114 can deposit saline into the tissue chamber 102. Many other examples of alternative locks or inlets are possible.
The drive shaft 104 can be an elongated rod at least partially housed by the tissue chamber 102 and extending vertically, or substantially vertically, through the tissue chamber 102. Additionally, in some embodiments, the drive shaft 104 includes a motor coupling 120 on a distal end 119 and the rotary blades 106 on a proximal end 121.
The motor coupling 120 can be coupled to a motor on an isolation chamber (shown in
Further, in some example embodiments, the drive shaft 104 includes a compression spring 118. The compression spring 118 can surround or substantially surround the drive shaft 104 and allow vertical movement of the rotary blades 106 along the drive shaft 104. The compression spring 118 can push the rotary blades 106 in position against the screen 112, while allowing the rotary blades 106 to adjust position along the drive shaft 104 and surpass potential blockages. Accordingly, large tissue chunks are progressively pushed through the openings of the screen 112, and the rotary blades 106 will not get stuck or stopped by large tissue chunks. In some embodiments, it is possible to adjust the compression force of that the rotary blades 106 apply to the tissue sample against the screen 112 with the spring tension adjustment nut 534 and lock nut 536 (as shown in
Additionally, in some examples, the drive shaft 104 and the rotary blades 106 can be configured to rotate in both clockwise and counter-clockwise directions.
The one or more rotary blades 106 are adjacent to the screen 112 and, in some examples, include stainless steel or another non-corrosive metal. Impeller forces of the rotating rotary blades 106 press the tissue sample through the screen 112 to process and break down the tissue sample into smaller pieces. In practice, rotation of the rotary blades 106 presses the tissue sample through the screen 112. Pressing the tissue sample through the screen 112, via the rotary blades 106, in this manner can be done in a sterile, full-immersion system to minimize tissue trauma.
In some examples, the tissue processing chamber 100 can include two rotary blades 106, as shown in
In some example embodiments, the rotary blades 106 can additionally be configured to pivot or rotate about a horizontal axis to facilitate various sizes, shapes, and consistency of different tissue samples.
The screen 112 can, for example, be a wire mesh. In some examples, the wire mesh can include non-corrosive metal, such as stainless steel, which is desirable, as the screen 112 must withstand repeated sterilization.
The screen 112 includes a plurality of pores 115 for the tissue sample to be pressed through. In some examples, pores 115 can be hexagonal (as shown in
Further, in some examples, the screen 112 can be removable and/or interchangeable such that the tissue processing chamber 100 can use a variety of different screens.
The support grid 108 is adjacent to and supports the screen 112. In some examples, the support grid 108 can also include pores 115. The pores of the support grid 108 can be larger than the pores of the screen 112. In practice, impeller forces of the rotating rotary blades 106 press the processed tissue through the support grid 108, in addition to the screen 112, to enter the collection chamber 110.
The collection chamber 110 is coupled to the tissue chamber 102 adjacent to the screen 112 and the support grid 108. In some examples, the collection chamber 110 can be conical. Many other shapes and configurations of the collection chamber 110 are possible.
Further, in example embodiments, the collection chamber 110 also includes an outlet 113 for outflow of processed tissue. The outlet 113 can attach to a sterile collection bag (not shown). Additionally or alternatively, the outlet 113 can attach to an outlet tube (as shown in
In some examples, the collection chamber 110 can be constructed of an autoclavable material (i.e., material that can withstand the pressure and temperature of tissue processing). For example, the collection chamber 110 can comprise a high grade polymer material. This is desirable, as tissue processing requires regulated temperatures and pressures.
Further, in some example embodiments, the tissue processing chamber 100 can include an O-ring seal 116 between the tissue chamber 102 and the collection chamber 110. In some examples, the O-ring seal 116 can create a static hermetic seal.
Additionally or alternatively, in some examples, the tissue chamber 100 can include two or more additional O-rings 109 and 111. These additional O-rings 109 and 111 can create a dynamic seal around the drive shaft 104.
In practice, specimen (e.g., a tissue sample) is deposited into the tissue chamber 102 via an inlet (e.g., the luer lock 114). A motor then spins the drive shaft 104 about a vertical (or substantially vertical) axis rotating the rotary blades 106. The rotating rotary blades 106 press the tissue through the through the screen 112, breaking down the processed tissue. Once the tissue passes through the screen 112, tissue collects in the collection chamber 110. In the example configuration shown in
In an alternate embodiment, the tissue processing chamber 100 is configured with the tissue chamber 102 below the collection chamber 110 (i.e., the tissue processing chamber 100 can be inverted or flipped upside down). This configuration is desirable in examples where the tissue sample includes a lipid. Namely, in practice, the lipids will float or rise to the top of the tissue chamber 102. Impeller forces of the rotary blades 106 will press the lipids through the screen 112 and into the collection chamber 110.
In some examples, the tissue processing chamber 100 can include a detachable stand 131. The detachable stand 131 can be configured to detachably fasten to the tissue chamber 102, as shown in
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The motor 324 can be configured to be on either the top or the bottom of the isolation chamber 322 to accommodate for different configurations of the tissue processing chamber 100. For example, as shown in
Additionally, portions of the tissue chamber 102, collection chamber 110, and/or O-ring seal 116 can couple to the isolation chamber 322. In some examples, the isolation chamber 322 can include locks 326 to stabilize the tissue processing chamber 100 during operation. It should be understood that any known type of connection mechanism can be used to attach the tissue processing chamber to the isolation chamber.
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Further, in some example embodiments, the tissue processing chamber 100 can include two rotary blades 106, as shown in
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Method 600 begins at block 602, which involves rotating at least one rotary blade within a tissue chamber.
At block 604, method 600 involves pressing at least a portion of a tissue sample through a screen adjacent to the at least one rotary blade via impeller forces of the at least one rotary blade. In some embodiments, before block 604, the method further involves depositing the tissue sample into the tissue chamber by way of a luer lock on the tissue chamber. Additionally, in some embodiments, the tissue chamber comprises a support grid adjacent to the screen and wherein the method further comprises pressing the processed tissue through the support grid.
At block 606, method 600 involves collecting processed tissue in a collection chamber. In some embodiments, method 600 further involves extracting the processed tissue from the collection chamber via a sterile collection bag attached to an outlet of the collection chamber.
Additionally, in some embodiments, method 600 can further involve depositing saline into the tissue chamber by way of a luer lock on the tissue chamber.
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While some embodiments have been illustrated and described in detail in the appended drawings and the foregoing description, such illustration and description are to be considered illustrative and not restrictive. Other variations to the disclosed embodiments can be understood and effected in practicing the claims, from a study of the drawings, the disclosure, and the appended claims. The mere fact that certain measures or features are recited in mutually different dependent claims does not indicate that a combination of these measures or features cannot be used. Any reference signs in the claims should not be construed as limiting the scope.
This application is a 35 U.S.C. 371 National Stage of International Application No. PCT/US21/25941 filed Apr. 6, 2021, which claims priority to U.S. Provisional Application No. 63/005,900, filed Apr. 6, 2020, the contents of which are hereby incorporated by reference in their entirety for all intents and purposes.
Filing Document | Filing Date | Country | Kind |
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PCT/US21/25941 | 4/6/2021 | WO |
Number | Date | Country | |
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63005900 | Apr 2020 | US |