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US 12,454,679 B2Granted
Non-decellularized plant leaf cultures for meat
AssigneeWorcester Polytechnic Institute
InventorsAlex S. Rebello, Jordan Jones, Glenn Gaudette
Life of this patent
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Abstract
An in vitro meat having non-decellularized leaf scaffold and a method for producing an in vitro meat using non-decellularized leaf scaffold are described herein. The method includes preparing a non-decellularized plant leaf and incubating the leaf in a cell culture media to obtain a non-decellularized leaf scaffold; seeding the leaf scaffold with a population of muscle cells to obtain a leaf scaffold adhered population of muscle cells; and growing the leaf scaffold adhered population of muscle cells in cell culture media thereby obtaining the in vitro meat.
Claims11
A method for producing an in vitro meat, the method comprising:
- preparing a non-decellularized plant leaf and incubating the leaf in a cell culture media to obtain a non-decellularized leaf scaffold formed from a spinach leaf;
- sterilizing the non-decellularized leaf scaffold with ethanol and a saline rinse;
- forming a population of muscle cells by:
- extracting a bovine tissue sample;
- incubating the sample in a digestion medium to form a tissue pellet;
- suspending the pellet in a growth medium to form satellite cells;
- and
- enriching the satellite cells by incubating in a carbon dioxide environment for removing unwanted cells to form a cell suspension including the population of muscle cells;
- seeding the non-decellularized leaf scaffold with the population of muscle cells to obtain a leaf scaffold adhered population of muscle cells, seeding further comprising:
- coating a cell culture plate with a hydrophobic polymer to obtain a polymer coated plate, the coating formed from polydimethylsiloxane (PDMS);
- placing the non-decellularized leaf scaffold on the polymer coated plate;
- laying a cloning well on the non-decellularized leaf scaffold;
- depositing the population of muscle cells on the non-decellularized leaf scaffold within the cloning well, the hydrophobic polymer and the cloning well maintaining the population of muscle cells in communication with the non-decellularized leaf scaffold;
- developing a muscle tissue layer on the leaf scaffold;
- assembling a plurality of the muscle tissue layers on the non-decellularized leaf scaffold for exposing the muscle cells to the cell culture media on opposed sides of the muscle tissue layers, further including layering a first leaf scaffold adhered population of muscle cells on a second leaf scaffold for forming an adhered population of muscle cells in a layered three-dimensional support structure of non-decellularized leaf scaffolds;
- removing non-adherent muscle cells from the non-decellularized leaf scaffold by washing the leaf scaffold with a buffer to obtain the leaf scaffold adhered population of muscle cells;
- and
- growing the leaf scaffold adhered population of muscle cells in cell culture media for differentiating the leaf scaffold adhered population of muscle cells in a differentiation media, and for achieving an alignment of a kappa value of between 0.17 to 0.45 for myosatellite cells defining the population of muscle cells, thereby obtaining the in vitro meat.
15 more elements
A method for producing an in vitro meat, the method comprising:
- providing at least one non-decellularized spinach plant leaf for a non-decellularized leaf scaffold;
- cleaning, cutting, sterilizing the non-decellularized leaf scaffold with ethanol and a saline rinse, and incubating the leaf in a cell culture media to obtain a non-decellularized leaf scaffold;
- forming a population of muscle cells by:
- extracting a bovine tissue sample;
- incubating the sample in a digestion medium to form a tissue pellet;
- suspending the pellet in a growth medium to form satellite cells;
- and
- enriching the satellite cells by incubating in a carbon dioxide environment for removing unwanted cells to form a cell suspension including the population of muscle cells;
- seeding the population of muscle cells on the non-decellularized leaf scaffold within a cloning well, seeding further comprising:
- placing the non-decellularized leaf scaffold on a polydimethylsiloxane (PDMS) coated plate;
- laying the cloning well on the non-decellularized leaf scaffold;
- depositing the population of muscle cells on the non-decellularized leaf scaffold within the cloning well, the cloning well maintaining the population of muscle cells in communication with the non-decellularized leaf scaffold from the hydrophobic PDMS;
- developing a muscle tissue layer on the leaf scaffold;
- assembling a plurality of the muscle tissue layers on the non-decellularized leaf scaffold for exposing the muscle cells to the cell culture media on opposed sides of the muscle tissue layers, further including layering a first leaf scaffold adhered population of muscle cells on a second leaf scaffold for forming an adhered population of muscle cells in a layered three-dimensional support structure of non-decellularized leaf scaffolds;
- removing non-adherent muscle cells from the non-decellularized leaf scaffold by washing the non-decellularized leaf scaffold with a buffer to obtain a leaf scaffold adhered population of muscle cells;
- and
- growing the leaf scaffold adhered population of muscle cells in cell culture media by differentiating the leaf scaffold adhered population of muscle cells in a differentiation media, for achieving an alignment of a kappa value of between 0.17 to 0.45 for myosatellite cells defining the population of muscle cells, thereby obtaining the in vitro meat.
14 more elements
Dependencies are inferred from the claim text.