Cell cultured berries and products derived therefrom

Abstract

The present invention provides culture-derived berry cells compositions which have a high phenolic acid and low anthocyanins in their polyphenolic fraction, and are high in protein, low in sugar, and have a high antioxidation potential.

Description

[0001] CELL CULTURED BERRIES AND PRODUCTS DERIVED THEREFROM

[0002] FIELD OF THE INVENTION

[0003] The present disclosure is generally directed to berry cell compositions. Specifically, the invention relates to berry cell culture-derived powder compositions.

[0004] BACKGROUND OF THE INVENTION

[0005] In the northern regions, including Scandinavian nations, Canada, the USA, Russia, and numerous EU member states, the consumption of berries is ingrained in daily dietary practices. The global berry market is projected to experience roughly 2% growth from 2020 to 2025. Berries are typically enjoyed in their fresh fruit form, but a diverse range of technological products featuring berries has gained substantial popularity in recent years. Particularly, berry-infused beverages and confectioneries have seen significant growth. This increased berry consumption can be attributed to a rising awareness of health consciousness and the availability of more processed "berry-packed" food options.

[0006] Strawberries Fragaria x ananas so). for example, have a long history of use worldwide. Both in fresh and processed forms, strawberries are the most popular berries in Europe, also thanks to their high nutritional and phytochemical contents, such as vitamins, anthocyanins, ellagitannins, quercetin, and catechin. Currently, strawberry consumption has attracted attention due to its biological properties, such as antioxidant, cardioprotective, anticancer, anti-inflammation, antidiabetic, antimetabolic syndrome, antiobesity, neuroprotective, and antimicrobial activities, related to human health benefits. For example, Human studies have suggested that regular strawberry ingestion increases endogenous antioxidant defenses.

[0007] Blackberries Rubus fruticosus) are also recognized for their antioxidant properties and are often promoted as a healthy addition to the diet. Blackberries and their by-products have been used since ancient times in traditional medicine, but recently the knowledge concerning their healthpromoting components has received a lot of attention, particularly due to their richness in different bioactive compounds, that mostly depend on the region, variety, time of harvest, soil composition, which, in turn, results in variations between cultivars produced in the same area.

[0008] Other berries, such as blueberries and chokeberries, are also known for their antioxidant properties and health benefits.

[0009] The health benefits associated with berries consumption are attributed to their content of phytochemicals, with anthocyanin pigments being particularly abundant. Among these phytochemicals, anthocyanins are believed to exert the most pronounced positive effects on health. Additional beneficial polyphenolic compounds include flavonoids (as quercetin, catechins, and myricetin), phenolic acids (gallic acid and ellagic acid), as well as other phytochemicals as vitamin C and E.

[0010] In the contemporary world, agriculture is undergoing significant transformations, primarily due to the impending requirement for escalated food production to cater to the expanding global population. Meeting this surging demand for sustenance on a global scale presents substantial environmental implications. Consequently, the predominant challenge of our era lies in simultaneously augmenting agricultural output while embracing and enacting sustainable production methodologies.

[0011] The cultivation of berries, like many crops, has some sustainability challenges, particularly concerning land use and environmental impact; for example, blueberries land use conversion is high and may disrupt local ecosystems, and the tendency toward monoculture farming may lead to reduced biodiversity, soil depletion, and increased vulnerability to diseases. The use of pesticides and fertilizers to maximize yields and the need for substantial irrigation (particularly in regions with dry climates) are yet additional challenges. All in all, berries have a high carbon footprint. This is mainly because of their high pesticide use, mechanized farming methods, refrigerated transportation, and use of plastic packaging. Addressing these sustainability issues in berries cultivation involves, among other practices, exploring alternative cultivation methods.

[0012] Plant cell suspension cultures offer the most reliable and productive system to generate natural products. Growing plant cells in vitro under controlled environmental conditions allows for precise control over cell growth and phytochemicals production and batch-to-batch product consistency. The use of plant cell cultures instead of whole plants allows products for the food, supplements, and cosmetics industries to be manufactured with less energy, lower possible impacts on the environment, and independent of location and season.

[0013] Considering the valuable health advantages offered by berries and the challenges associated with their cultivation, there is a need to establish a sustainable in vitro cell culture system for berries.

[0014] SUMMARY OF THE INVENTION

[0015] The following embodiments are described and illustrated in conjunction with compositions and methods which are meant to be exemplary and illustrative, not limiting in scope. In various embodiments, one or more of the above-described problems have been reduced or eliminated, while other embodiments are directed to other advantages or improvements.

[0016] In some embodiments, there is provided composition including culture-derived berry cells, wherein the culture-derived berry cells contain a polyphenolic fraction including at least about 50% phenolic acids.

[0017] In some embodiments, the culture-derived berry cells include at least about 0.5%, 0.6%, 0.7%, or 0.8% w / w polyphenols of dry cells weight. In some embodiments, the culture-derived berry cells include at least about 12% w / w protein of dry cells weight. In some embodiments, the culture-derived berry cells include less than about 20% w / w sugar of dry cells weight. In some embodiments, the culture-derived berry cells include at least about 20% w / w dietary fibers of dry cells weight.

[0018] To clarify, the term “dry cell weight” means the weight of the dried culture-derived berry cells in the composition.

[0019] In some embodiments, the culture-derived berry cells include a secondary metabolites fraction including at least about 30% polyphenols. In some embodiments, the culture-derived berry cells include a secondary metabolites fraction including at least about 25% phenolic acids. In some embodiments, the culture-derived berry cells include a secondary metabolites fraction including at least about 20% terpenoids. In some embodiments, the culture-derived berry cells include a secondary metabolites fraction including at least about 10% alkaloids. In some embodiments, the % is a molecular percentage (% of the specific molecules out of the total number of molecules). In some embodiments, the % is % w / w. In some embodiments, the % is determined by mass spectrometry analysis.

[0020] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes at least about 60% non-flavonoids polyphenols. In some embodiments, the polyphenols fraction of the culture-derived berry cells includes at least about 80% phenolic acids. In some embodiments, the polyphenols fraction of the culture-derived berry cells includes less than about 10% flavonoids. In some embodiments, the polyphenols fraction of the culture-derived berry cells includes less than about 5% anthocyanins. In some embodiments, the polyphenols fraction of the culture-derived berry cells includes less than about 5% tannins. In some embodiments, the polyphenols fraction of the culture-derived berry cells includes at least about 1% lignans and coumarins. In some embodiments, the % is a molecular percentage (% of the specific molecules out of the total number of molecules). In some embodiments, the % is % w / w. In some embodiments, the % is determined by mass spectrometry analysis.

[0021] In some embodiments, the composition is in the form of a dry powder. In some embodiments, the dry powder is a freeze-dried, a spray dried, or a fluid bed dried powder. In some embodiments, the composition is in the form of a liquid suspension.

[0022] In some embodiments, the composition includes at least about 70%, 80%, or 90% w / w whole berry cells. In some embodiments, the composition consists essentially of berry culture-derived whole cells.

[0023] In some embodiments, the composition is edible.

[0024] In some embodiments, the composition is not sweet.

[0025] In some embodiments, the berry cells are selected from strawberry cells, blackberry cells, blueberry cells, chokeberry cells, elderberry cells, raspberry cells, and a combination thereof. In some embodiments, the berry cells are from a genus selected from Fragaria, Rubus, Vaccinium, Aronia, Sambucus, and a combination thereof. In some embodiments, the berry cells are from Fragaria x ananassa and / or Rubus Fruticosus.

[0026] In some embodiments, the composition further includes a cell culture medium. In some embodiments, the medium includes a hormone, 2-Morpholinoethanesulfonic acid monohydrate (MES), and / or amino acids.

[0027] In some embodiments, there is provided a food product including the berry cells composition disclosed herein.

[0028] In some embodiments, there is provided a food supplement including the berry cells composition disclosed herein.

[0029] In some embodiments, there is provided a nutraceutical including the berry cells composition disclosed herein.

[0030] In addition to the exemplary embodiments described above, further embodiments will become apparent by reference to the figures and by study of the following detailed descriptions.

[0031] BRIEF DESCRIPTION OF DRAWINGS

[0032] The invention will now be described in relation to certain examples and embodiments with reference to the following illustrative figures.

[0033] Fig. 1 shows strawberries cell culture growth rate including total dissolved solids (TDS) and Brix follow-up.

[0034] Fig. 2 shows blackberries cell culture growth rate including TDS and Brix follow-up.

[0035] Figs. 3A-3B show secondary metabolites content in strawberry (Fig. 3A) and blackberry (Fig. 3B) cell cultures. Percentages are based on mass spectrometry analysis.

[0036] Fig. 4 shows an acellular Reactive Oxygen Species (ROS) scavenging capacity of a polyphenols extract of a strawberry cell culture (right) compared to a polyphenol extract from fresh strawberry fruits (left), evaluated by Oxygen Radical Absorbance Capacity (ORAC) assay. PP: polyphenols; TE: Trolox Equivalents.

[0037] Figs. 5A-5B show cell viability upon treatment with polyphenolic extracts of strawberry fruit and cell. HaCaT cells were treated with the specified samples, and cell viability was measured by an MTT assay. Fig. 5A. Fruit polyphenol extract; Fig. 5B. Cell culture polyphenol extract.

[0038] Figs. 6A-6B shows cellular antioxidant capacity. ROS scavenging capacity of the polyphenolic extracts was evaluated by the deFDA method. Results are presented as means of percent of inhibition ± SEM, n=3. Fig. 6A. fruit polyphenolic extract; Fig. 6B. Cell culture polyphenol extract. NAC, N- acetylcysteine (positive control).

[0039] DETAILED DESCRIPTION OF THE INVENTION

[0040] In the following description, various aspects of the disclosure will be described. For the purpose of explanation, specific configurations and details are set forth in order to provide a thorough understanding of the different aspects of the disclosure. However, it will also be apparent to one skilled in the art that the disclosure may be practiced without specific details being presented herein. Furthermore, well-known features may be omitted or simplified in order not to obscure the disclosure.

[0041] The present invention provides cell culture products derived from berry cells.

[0042] These berry cells may originate from various parts of the berry plant, including edible components such as leaves or fruit, as well as stem segments. The cultured cells contain certain desirable features, such as a high content of proteins and low content of sugars, in addition to a high antioxidation potential and high level of phenolic acids, together with low levels of flavonoids and anthocyanins. Phenolic acids are assumed to have higher bioavailability than anthocyanins, as they are smaller compounds. In addition, our results show that our specific composition of polyphenols is more potent in reducing Reactive Oxygen Species (ROS) than polyphenol extract from berry fruit, which is primarily composed of anthocyanins. In addition, phenolic acids have stronger antimicrobial properties than anthocyanins.

[0043] The products of the invention is valuable for various applications in the food, pharmaceutical, and nutraceutical industries. The invention provides a novel and sustainable source of berry-derived antioxidants and polyphenols. This cell culture system allows the production of these bioactive compounds on a larger scale, independent of seasonal variations and geographic limitations.

[0044] Culture-derived berry cell compositions

[0045] In some embodiments, there is provided a composition including culture-derived berry cells, wherein the culture-derived berry cells contain a polyphenolic fraction including at least about 50% phenolic acids. As can be seen from Table 3 below, the claimed culture-derived berry cells compositions advantageously include a polyphenolic fraction with a different composition compared to that of berry fruit. Some of the most distinctive differences in the polyphenolic fraction composition are a much higher content of phenolic acids (about 90% compared to less than 40% in fruit) and a much lower content of anthocyanins (less than 0.5% compared to more than 20% in fruit). As noted above, the smaller size of phenolic acids compared to anthocyanins are expected to provide for a higher bioavailability. Furthermore, as can be seen from Figs. 4 and 6, the antioxidant potential of the composition of the invention is much higher than that of fruit.

[0046] The term “berry” or “berries” refers to a small, pulpy, and edible fruit, typically juicy, rounded, and may contain pips or seeds. The seeds are typically small and embedded within the fruit rather than having a large stone or pit. Berries can range in taste from sweet to tart and may refer to, including but not limited to, blueberries, chokeberries, blackberries, boysenberries, brambles, cranberries, currants (including red, white, and black currants), elderberries, gooseberries, huckleberries, loganberries, marionberries, mulberries, prairie berries, raspberries, saskatoon berries, strawberries, tayberries, and the like or any combination thereof. The composition may include a single species of berries, or a combination of several species. Additionally, the berry cells may be derived from a single plant part or from a combination of plant parts.

[0047] In some embodiments, the berries from which the culture is derived are natural berries. In some embodiments, the berries from which the culture is derived are genetically modified berries.

[0048] The term "plant part" refers to a specific component or portion of a plant, which can be either a distinct anatomical structure or a functional part of the plant, including but not limited to leaves, stems, roots, flowers, fruits, seeds, tendrils, thorns, bulbs, hypocotyl, cotyledon, or any combination thereof. The plant part may be a mature plant part or an embryonic plant part.

[0049] In some embodiments, the berry plant part is selected from fruit, leaf, stem, seed, and a combination thereof.

[0050] The composition may be in any suitable form, such as in liquid form or in a dry form.

[0051] In some embodiments, the composition is in the form of a liquid suspension. The cultured berry cells may be suspended in any suitable liquid, such as, but not limited to beverages such as a juice or a shake and other liquid food products. The composition may also be used as a wet biomass, following filtration of media, as a food supplement.

[0052] In some embodiments, the composition includes culture-derived berry cells suspended in a culture medium.

[0053] In some embodiments, the culture medium includes a synthetic hormone. In some embodiments, the culture medium includes a hormone selected from a cytokinin and an auxin. In some embodiments, the hormone is selected from zeatin, meta topolin, benzyl adenine (BA), kinetin, thidiazuron (TDZ), adenine hemisulfate (AdS), 1 -Naphthaleneacetic acid (NAA), Indole - 3-acetic acid (IAA), indole-3 -butyric acid (IBA), 2, 4 -dichlorophenoxy acetic acid (2,4-D), and combinations thereof. In some embodiments, the hormone is selected from meta topolin, benzyl adenine (BA), kinetin, thidiazuron (TDZ), adenine hemisulfate (AdS), 1 -Naphthaleneacetic acid (NAA), indole-3 -butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), and combinations thereof.

[0054] In some embodiments, the culture medium includes 2-Morpholinoethanesulfonic acid monohydrate (MES).

[0055] In some embodiments, the culture medium includes amino acids.

[0056] In some embodiments, the composition is in the form of a dry powder. The composition may be dried by any suitable way, such as by lyophilizing, freeze-drying, a spray drying, etc. In some embodiments, the dry powder is a freeze-dried, a spray dried, or a fluid bed dried, powder.

[0057] In some embodiments, the culture-derived berry cells include at least about 0.5%, 0.6%, 0.7%, or 0.8% w / w polyphenols of the cells dry weight. In some embodiments, the culture-derived berry cells include about 0.5%-10%, 0.8%-5%, or l%-4% w / w polyphenols of the cells dry weight. In some embodiments, the culture-derived berry cells include at least about 0.5%, 0.8%, 1%, 2%, 3%, or 4% w / w polyphenols of the cells dry weight. In some embodiments, the culture-derived berry cells include less than about 20%, 15%, 10%, 9%, 8%, 7%, 6% or 5% w / w polyphenols of the cells dry weight.

[0058] In some embodiments, the culture-derived berry cells include at least about 10%, 12%, 15%, or 20% w / w protein of the cells dry weight. In some embodiments, the culture-derived berry cells include between about 10%-30%, 12%-30%, 15%-30%, 18%-26%, or 20%-26% w / w protein of the cells dry weight.

[0059] In some embodiments, the composition includes less than about 40%, 35%, 30%, 25%, or 20% w / w sugar of the cells dry weight. In some embodiments, the culture-derived berry cells include about 5%-35%, 5%-30%, 10%-30%, 10%-25%, or 10%-20% w / w sugar of the cells dry weight.

[0060] In some embodiments, the culture-derived berry cells include at least about 15%, 20%, or 25% w / w dietary fibers of the cells dry weight. In some embodiments, the culture-derived berry cells include between about 15%-35% or 20%-30% w / w dietary fibers of the cells dry weight. Plant secondary metabolites are organic compounds produced by plants that are not directly involved in primary growth, development, or reproduction. Unlike primary metabolites (such as sugars, amino acids, and lipids), which are essential for basic cellular functions, secondary metabolites often have multiple functions such as the regulation of plant growth and development, participation in plant innate immunity, and response to environmental stresses. Plant secondary metabolites are mainly classified into molecular families based on their biosynthesis pathways: phenolics (including phenolic acids, tannins, flavonoids, anthocyanins, etc.), terpenes, steroids, and alkaloids.

[0061] It is noted that the secondary metabolites fraction of ripe berries typically contains about VO- 95 % polyphenols, about 5-15% terpenoids, and about 5-10% alkaloids.

[0062] In some embodiments, the secondary metabolites fraction of the culture-derived berry cells includes at least about 25%, 30%, 32%, 34%, 36%, 38%, or 40% polyphenols.

[0063] In some embodiments, the secondary metabolites fraction of the culture-derived berry cells includes at least about 25%, 28%, 30%, 32%, 34%, 36%, 38%, or 40% phenolic acids.

[0064] In some embodiments, the secondary metabolites fraction of the culture-derived berry cells includes at least about 15%, 18%, 20%, 22%, 25%, 28%, or 30% terpenoids.

[0065] In some embodiments, the secondary metabolites fraction of the culture-derived berry cells includes at least about 10%, 15%, 17%, 20%, 22%, or 24% alkaloids.

[0066] In some embodiments, the % is a molecular percentage (% of the specific molecules out of the total number of molecules). In some embodiments, the % is % w / w. In some embodiments, the % is determined by mass spectrometry analysis.

[0067] The term “secondary metabolite fraction”, as used herein, relates to the total amount of secondary metabolites present in the culture-derived berry cells.

[0068] In some embodiments, the ratio of polyphenols to terpenoids is at least about 1:1, 1.2:1, or 1.5:1.

[0069] In some embodiments, the ratio of polyphenols to alkaloids is at least about 1:1, 1.2:1, 1.5:1, or 2:1.

[0070] In some embodiments, the ratio of polyphenols to terpenoids, and / or alkaloids is in the culture-derived berry cells. In some embodiments, the ratio of polyphenols to terpenoids, and / or alkaloids is in the polyphenolic fraction of the culture-derived berry cells.

[0071] The term “polyphenolic fraction”, as used herein, relates to the total amount of polyphenolic molecules present in the culture-derived berry cells. The polyphenolic fraction is usually analyzed following extraction with a combination of solvents such as alcohol and water at varying ratios.

[0072] The polyphenolic fraction of the culture-derived berry cells may be extracted for analysis by any suitable method known in the art, including, but not limited to, solvents such as methanol, ethanol, and acetone. A non-limiting example is by 60-80% methanol extraction, centrifugation at about 6000 rpm and filtering through a 0.22 micron hydrophobic polytetrafluoroethylene (PTFE) syringe filter, (see Example 3). It should be clarified that following extraction, the polyphenolic fraction is not 100% polyphenols, but includes a certain % of polyphenols, which is considered to be the total (100%) for calculating the composition of the polyphenolic fraction.

[0073] Identification of compounds may also be carried out by any suitable method and depending on the compounds to be identified. Examples for such methods include, but are not limited to, mass spectrometry (e.g., MS-MS, LC-MS, GC-MS), liquid chromatography (HPLC), etc.

[0074] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes at least about 50%, 60%, 70%, 80%, 90%, or 95% non-flavonoids polyphenols.

[0075] In some embodiments, the non-flavonoids polyphenols of the polyphenolic fraction of the culture-derived berry cells include at least about 60%, 70%, 80%, 90%, or 95% phenolic acids.

[0076] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes at least about 50%, 60%, 70%, 80%, 85%, 90%, or 95% phenolic acids.

[0077] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes less than about 30%, 25%, 20%, 15%, 10%, 8%, 7%, 6%, 5%, 4%, or 3% flavonoids.

[0078] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes less than about 20%, 15%, 10%, 5%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% or anthocyanins. In some embodiments, the polyphenolic fraction of the culture-derived berry cells essentially lacks anthocyanins.

[0079] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes less than about 5%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% tannins. In some embodiments, the polyphenolic fraction of the culture-derived berry cells essentially lacks tannins.

[0080] In some embodiments, the polyphenolic fraction of the culture-derived berry cells includes at least about 1%, 2%, 3%, 4%, 5%, or 6% lignans and coumarins.

[0081] In some embodiments, the % is a molecular percentage (% of the specific molecules out of the total number of molecules). In some embodiments, the % is % w / w. In some embodiments, the % is determined by mass spectrometry analysis. In some embodiments, the ratio of phenolic acids to anthocyanins is at least about 100:1, 150:1, 180:1, 200:1, 250:1, 300:1, 350:1, 400:1, 500:1, 1,000:1, 2,000:1, 5,000:1, or 10,000:1. In some embodiments, the ratio of phenolic acids to anthocyanins is at least about 220:1.

[0082] In some embodiments, the ratio of phenolic acids to tannins is at least about 100:1, 150:1, 180:1, 200:1, 250:1, 300:1, 350:1, 400:1, 500:1, 1,000:1, 2,000:1, 5,000:1, or 10,000:1.

[0083] In some embodiments, the ratio of phenolic acids to flavonoids is at least about 5:1, 10:1, 15:1, 20:1, 30:1, 40:1, or 50:1.

[0084] In some embodiments, the ratio of phenolic acids to lignans and coumarins is at least about 5:1, 10:1, 20:1, 30:1, or 40:1.

[0085] In some embodiments, the ratio of phenolic acids to anthocyanins, tannins, flavonoids, and / or lignans and coumarins is in the culture-derived berry cells. In some embodiments, the ratio of phenolic acids to anthocyanins, tannins, flavonoids, and / or lignans and coumarins is in the polyphenolic fraction of the culture-derived berry cells.

[0086] The term "essentially lacks" means that the composition may have omitted, substantially absent, or largely reduced additional ingredients, but only if the absent ingredients do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

[0087] In some embodiments, the culture-derived berry cells include at least about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 0.95%, 1%, 2%, or 4% non-flavonoids polyphenols per dry weight.

[0088] In some embodiments, the culture-derived berry cells include at least about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 0.95%, 1%, 2%, or 4% phenolic acids per dry weight.

[0089] In some embodiments, the culture-derived berry cells include less than about 0.8%, 05%, 0.2%, 0.15%, 0.1%, 0.05%, 0.01%, or 0.005% anthocyanins per dry weight.

[0090] In some embodiments, the culture-derived berry cells include less than about 0.2%, 0.1%, 0.05%, 0.01%, or 0.005% tannins per dry weight.

[0091] In some embodiments, the culture-derived berry cells include less than about 1%, 0.5%, 0.3%, 0.25%, 0.2%, 0.15%, or 0.1% flavonoids per dry weight.

[0092] In some embodiments, the culture-derived berry cells include at least about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.1%, 0.15%, or 0.2% lignans and coumarins per dry weight.

[0093] In some embodiments, the culture-derived berry cells essentially lack anthocyanins. In some embodiments, the culture-derived berry cells essentially lack tannins.

[0094] In some embodiments, the % is a molecular percentage (% of the specific molecules out of the total number of molecules). In some embodiments, the % is w / w %. In some embodiments, the % is determined by mass spectrometry analysis.

[0095] It is appreciated that the percentages provided for compositions of the secondary metabolites fraction and of the polyphenolic fraction are based on mass spectrometry analysis. Accordingly, these percentages may generally correspond to molecular %, namely the % of the specific molecules out of the total.

[0096] In some embodiments, the berry cells composition includes about 60-100%, 70-99%, 75- 99%, 80-99%, or 85-99% w / w berry culture-derived whole cells. In some embodiments, the berry cells composition includes at least about 60%, 70%, 75%, 80%, 85%, or 90% w / w culture-derived whole berry cells.

[0097] The term "whole cells" refers to cells that are generally intact and may become functional in a suitable environment. The whole cells include all cellular components and structures, such as cell wall, cytoplasm, and various organelles, including but not limited to, the nucleus, mitochondria, endoplasmic reticulum, and any other cellular structures that actively contribute to the form and function of the cell. It is noted that the cell wall may have increased porosity compared to the original plant cell.

[0098] In some embodiments, the composition consists essentially of culture-derived whole berry cells.

[0099] The term "consisting essentially of" means that the composition may include additional ingredients, but only if the additional ingredients are included in minute amounts and do not materially alter the basic and novel characteristics of the claimed composition.

[0100] In some embodiments, the composition is edible. In some embodiments, the composition is not sweet.

[0101] In some embodiments, the berry cells are selected from strawberry cells, blackberry cells, blueberry cells, chokeberry cells, elderberry cells, raspberry cells, cells and a combination thereof.

[0102] In some embodiments, the berry cells are from a genus selected from Fragaria, Rubus, Vaccinium, Aronia, Sambucus, and a combination thereof.

[0103] In some embodiments, the berry cells are from Fragaria x ananassa and / or Rubus fruticosus.

[0104] In some embodiments, there is provided a food product including the berry cells composition disclosed herein. In some embodiments, the food product including the berry cells composition can be in the form of juice, beverage, sauce, dressing, soup, shake, yogurt, smoothies, etc.

[0105] In some embodiments, there is provided a food supplement including the berry cells composition disclosed herein. In some embodiments, there is provided a nutraceutical composition including the berry cells composition disclosed herein.

[0106] The term “food supplement”, as used herein, relates to a product includes one or more nutrients, such as vitamins, minerals, herbs, amino acids, enzymes, or other bioactive substances, in addition to the berry cell composition, in order to supplement the diet. The supplements may be in any suitable form, preferably for oral administration, such as capsules, tablets, powders, or liquids.

[0107] The term “nutraceutical composition”, as used herein, relates to a formulation that combines nutrients (such as vitamins, minerals, amino acids, or fatty acids) with the berry cell composition, so as to provide health benefits beyond basic nutrition. The nutraceutical composition may be in any relevant shape or form, including capsules, tablets, powders, or liquids, and may include antioxidants, probiotics, herbal extracts, or omega-3 fatty acids.

[0108] Methods for preparing the culture-derived berry cells composition

[0109] The culture-derived berry cells compositions may be prepared by any suitable method of cell culture. Provided herein and in the Examples are some nonlimiting exemplary methods for preparing the culture-derived berry cells compositions of the invention.

[0110] In some embodiments, there is provided a method for preparing the culture-derived berry cells compositions of the invention, the method including the steps of: an explant acclimatization step; a callus formation step; a callus proliferation step; and a cell suspension establishment step.

[0111] Definitions and embodiments mentioned above and which may be relevant to the present embodiments also apply here, and vice versa. Some particularly relevant embodiments may be pointed out or explicitly repeated. For terms used herein, unless stated otherwise, their definition and embodiments are intended to be the same as above (mill al is mutandis).

[0112] In some embodiments, the explant acclimatization step includes: removing an explant from a berry plant and putting the explant in a semi-solid induction medium containing ingredients selected from Table 1 (including at least one carbon source) and further including at least one gelling agent, keeping the explant in the dark; and dissecting the explant into sections and placing the sections in plates containing a medium including ingredients selected from Table 1, and adding at least one antioxidant and a phenols absorber, to obtain induced explant sections.

[0113] In some embodiments, the at least one gelling agent is selected from agar, Gelrite™, and combinations thereof.

[0114] In some embodiments, the at least one carbon source is selected from sucrose, glucose, maltose, fructose, and combinations thereof.

[0115] In some embodiments, the at least one antioxidant is selected from ascorbic acid, citric acid, dithiothreitol (DTT), and combinations thereof.

[0116] In some embodiments, the phenols absorber is polyvinyl pyrrolidone.

[0117] In some embodiments, the callus formation step includes: putting the induced explant sections in a semi-solid induction medium containing ingredients selected from Table 1 (including a carbon source) and further including at least one gelling agent but without antioxidants and phenols absorber; adding at least one auxin; and incubating for several days to obtain a formed callus.

[0118] In some embodiments, at least one auxin is selected from 1 -Naphthaleneacetic acid (NAA), Indole-3-acetic acid (IAA), indole- 3 -butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), and combinations thereof.

[0119] In some embodiments, the callus proliferation step includes: adding to the formed callus at least one cytokinin; incubating until growth is observed; and adding at least one amino acid source to the callus, to obtain a friable callus.

[0120] In some embodiments, the at least one cytokinin is selected from zeatin, meta topolin, benzyl adenine (BA), kinetin, thidiazuron (TDZ), adenine hemisulfate (AdS), and combinations thereof.

[0121] In some embodiments, the at least one amino acid source is selected from casein hydrolysate, yeast extract, free amino acids, and combinations thereof.

[0122] In some embodiments, the cell suspension establishment step includes: adding liquid medium containing ingredients from Table 1 (but no gelling agents) and 2- Morpholinoethanesulfonic acid monohydrate (MES) buffer to the friable callus, and incubating to establish cell suspension.

[0123] In some embodiments, the liquid medium includes a higher content of phosphorus sources such as KH2PO4, (NH4)H2PO4, and NaH2PO4Compared to the semi-solid medium. In some embodiments, the culture-derived berry cells composition is further filtered to remove the medium and isolate intact culture-derived berry cells. In some embodiments, the culture-derived berry cells include is dried, as described above.

[0124] In some embodiments, there is provided a culture-derived berry cells composition prepared by the methods described herein.

[0125] In some embodiments, culture-derived berry cells composition is prepared by a method which includes a step including adding an amino acid source.

[0126] In some embodiments, culture-derived berry cells composition is prepared by a method which includes a step including adding MES.

[0127] In some embodiments, culture-derived berry cells composition is prepared by a method which includes a step including adding a hormone. In some embodiments, the hormone is selected from an auxin and a cytokinin. In some embodiments, the hormone is a synthetic hormone. In some embodiments, the hormone is selected from zeatin, meta topolin, benzyl adenine (BA), kinetin, thidiazuron (TDZ), adenine hemisulfate (AdS), 1 -Naphthaleneacetic acid (NAA), Indole - 3-acetic acid (IAA), indole-3 -butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), and combinations thereof. In some embodiments, the hormone is selected from meta topolin, benzyl adenine (BA), kinetin, thidiazuron (TDZ), adenine hemisulfate (AdS), 1 -Naphthaleneacetic acid (NAA), indole-3 -butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), and combinations thereof.

[0128] In some embodiments, culture-derived berry cells composition is prepared by a method not including adding coconut water.

[0129] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[0130] The term "a" and "an" refers to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0131] The term "about" when referring to a measurable value such as an amount, a ratio, and the like, is meant to encompass variations of ±10% of the indicated value, as such variations are also suitable to perform the disclosed invention. Any numerical values appearing in the application are intended to be construed as if preceded by “about”, unless indicated otherwise.

[0132] While certain embodiments of the invention have been illustrated and described, it will be clear that the invention is not limited to the embodiments described herein. Numerous modifications, changes, variations, substitutions, and equivalents will be apparent to those skilled in the art without departing from the spirit and scope of the present invention as described by the claims, which follow.

[0133] The following examples are presented in order to more fully illustrate some embodiments of the invention. They should in no way be construed, however, as limiting the broad scope of the invention. One skilled in the art can readily devise many variations and modifications of the principles disclosed herein without departing from the scope of the invention.

[0134] EXAMPLES

[0135] Example 1: Callus establishment from strawberries and blackberries

[0136] Plant material sterilization

[0137] Whole berries were rinsed to surface sterilize in running water for 30 minutes, then washed with 1% spore kill (v / v) solution for 2 minutes. The berries were rinsed in 70% ethanol for 1 minute and were washed in 2% sodium hypochlorite (w / v) with 0.02% (v / v) Tween 20 for 15 minutes followed by 5 rinses in sterile distilled water.

[0138] Explants acclimation

[0139] Explants from the plant material were dissected in sterile environment into 3-8 mm sections consisting of berry skin and flesh and placed on a semi-solid culture media with the flesh side down, leaving only the skin exposed to the air, in order to prevent the flesh tissue from oxidation and furthermore cell necrosis. Explant sections were placed in 94x16mm culture plates containing 25 ml of autoclaved media such as Gamborg's B5, Anderson basal salt mixture, Schenk & Hildebrandt Basal salt medium, Chee & Pool, and Woody plant media (WPM), containing ingredients selected from Table 1 (including a carbon source such as sucrose, glucose, maltose, fructose, or their combination) at the indicated concentration ranges, with the addition of gelling agents.

[0140] For the explants acclimation stage, media also included antioxidants agents that were filter sterilized and added post autoclave, as well as phenols absorber (see Table 1).A11 media were sterilized by autoclaving for 25 minutes at 121° C and 118 Kpa. The plates were kept in the dark at 25°C

[0141] Callus formation

[0142] For callus formation the explants are subcultured in a semi-solid medium containing ingredients selected from Table 1 (including a carbon source) at the indicated concentration ranges, with the addition of gelling agents, but excluding the antioxidants and phenols absorbers, and with the addition of auxins (see Table 1). First signs of callus formation were seen after 7-10 days around the explants girth. Callus was isolated from the explant and sub-cultured every 3-4 weeks.

[0143] Callus proliferation

[0144] For callus proliferation, cytokinins (selected from Table 1) were added. The derived callus showed rapid growth rate. For friable callus formation, amino acid sources as casein hydrolysate or yeast extract as well as free amino acids were added to the media. Friable cell lines were selected for further maintenance.

[0145] Table 1: Media composition ingredients for berry cells

[0146] In bold: required ingredients - CuSCL, FeNaEDTA, H3BO3, MnSCL, Na2MoO4, ZnSCL, MgSCL, myo-inositol, Bl, DTT, PVP 10, zeatin, kinetin, TDZ, casein hydrolysate, 2,4-D, and a carbon source.

[0147] Example 2: Cell suspension establishment

[0148] Established friable cell lines derived from semi-solid media were chosen for initiation of liquid suspensions. Cell Suspensions were created by introducing 2 grams of callus into 20 ml liquid medium in 100 ml sterile Erlenmeyer flasks. The liquid medium used for the suspension establishment was based on the semi-solid medium composition with minor modification such as higher concentrations of phosphorus sources such as KH2PO4, (NJL EhPC and NatEPCU, and lacking a gelling agent. In addition, MES buffer was added for stabilizing the pH values of the suspension. The flasks were covered with sterile aluminum foil and agitated at 110 revolutions per minute (rpm) in orbital shaker. The suspensions were kept in darkness at 25°C. Cultures were routinely sub-cultured every 7-10 days to fresh growing medium, the spent media was filtered, and fresh medium was added. The cells growth was determined by measuring fresh and dry weight biomass. In addition, total dissolved solids (TDS) and Brix (measurement of soluble solids concentration, primarily sugars) were measured over time. TDS is generally used for monitoring cell growth, and is usually inversely correlated to cell biomass. Brix is used in order to monitor sugar content in the medium, and as cell grow- they consume more sugar and the Brix value decreases.

[0149] As can be seen from the figures, the strawberry cell culture presented growth rate of 10.24 grams in 14 days (Fig. 1), and the blackberry cell culture presented growth rate of 7.44 grams in 9 days (Fig. 2). Both cell cultures exhibited standard TDS and Brix values through the growth cycles.

[0150] Example 3: Analysis of total polyphenol content in cell cultures

[0151] Preparation and extraction of phenolic compounds (polyphenol extract) were made from various berry cell lines obtained from liquid cultures. The cells were washed with distilled water several times, filtered through a 500-micron cell strainer, dried on Whatman filter paper, transferred to centrifuge tubes, and frozen with liquid nitrogen for a further freeze drying (lyophilizing). Samples were dried for 48h. Dried cells were ground to obtain a fine powder.

[0152] Phenolic compounds were extracted using 60-80% methanol solution in ratio of 10:1 solvent to sample. Samples were shaken for 10 min at 1200 rpm at room temperature, and the phenolic fraction separated from the cells using centrifuge at 6000 rpm and filtered through 0.22 micron hydrophobic polytetrafluoroethylene (PTFE) syringe filter. Samples were stored at -20°C until use.

[0153] Phenolic compounds content was determined in triplicate by the Folin-Ciocalteu spectrophotometric method (Singleton & Rossi, 1965, American Journal of Enology and Viticulture, 16, 144-158) with some modifications. A lOOul aliquot from each extract was added to 125ul Folin-Ciocalteu reagent solution and 1.5ml distilled water. The solution was left to rest for 5 min before being added with 375ul of 20% sodium carbonate solution, and then vortexed and left to rest for 10 min. Distilled water (475ul) was added to the solution and vortexed, and the resulting color absorbance was measured at 765 nm in a UV-Vis spectrophotometer. To obtain the phenolic compounds’ concentration data, a calibration curve was constructed using different concentrations of gallic acid (50 to 500 ug / ml). Total phenolic compounds’ content was expressed in mg of gallic acid equivalent (GAE) per g of sample (based on dry matter).

[0154] The total polyphenolic content of strawberry cells was 0.8-3.5% (based on GAE) (w / w), and that of blackberry cells was 1.5-4% (based on GAE) (w / w).

[0155] Example 4: Nutritional composition of the strawberry cell culture

[0156] For nutritional composition analysis, strawberry cell cultures were washed three times with distilled water, filtered through a 100 micron mesh, dried, and kept at -20°C prior to use.

[0157] The percentages of total protein, total fat (by hydrolysis), total carbohydrates (calculated), sugar, ash, and moisture (% water content) were determined by standard methods, and are presented in Table 2.

[0158] In addition, a comparison of the nutritional values of the strawberry cell cultures of the invention to known strawberry fruit powder (e.g., see the US department of agriculture publication in “FoodData Central” at https: / / fdc.nal.usda.gov / food-details / 2346409 / nutrients) is also presented in Table 2.

[0159] Table 2: Composition analysis of dried cultured strawberry cells vs. strawberry fruits (w / w%) As can be seen from Table 2, the dried strawberry cultured cells of the invention include a lower sugar content and a higher protein content compared to dried fruit.

[0160] Example 5: Metabolomic profiling of strawberry cell culture

[0161] For identification of secondary metabolites, two samples of strawberry cell cultures and two samples of blackberry cell cultures were freeze-dried before being ground into powder by a grinder for 90 s at 30 Hz. Then, 100 mg powder of each sample was dissolved in 1.2 mL of 70% methanol extract. The mixture was vortexed for 30 s and placed 30 min. This operation was repeated 6 times. Then, the mixture of each sample was placed in a refrigerator at 4 °C overnight. Next, each mixture was centrifuged for 10 min at 11,304 x g and the supernatant was filtered through a 0.22 pm pore size microporous membrane. The filtrate was transferred into a detection bottle and used for UPLC-MS / MS analysis. The analyzer system included Ultra Performance Liquid Chromatography (UPLC) (SHIMADZU Nexera X2, Kyoto, Japan) and Tandem Mass Spectrometry (MS / MS) (Applied Biosystems 4500 QTRAP®, AB Sciex, Framingham, MA, USA). Qualitative analysis of the compounds was performed based on a self-construction database and quantitative analysis relied on multiple reaction monitoring (MRM) mode. Analyst 1.6.3 (https: / / sciex.com / products / software / analystsoftware) was used for processing the mass spectrometry data. Results of secondary metabolites distribution in strawberry and blackberry cell samples are presented in Figs. 3A-3B. In both samples, total phenolics represented the majority of identified secondary metabolites. The two other major groups identified were alkaloids and terpenoids.

[0162] Typically, dried berries fruit powders contain high concentrations of flavonoids, most of which are anthocyanin-type (expressed in the intense fruit color). In contrast, as shown in Table 3, the strawberry and blackberry cell cultures prepared according to the invention contain a different composition of polyphenols, with a lower concentration of flavonoids and anthocyanins, relative to dried fruit powders. Table 3 summarize the distribution of polyphenols classes relative to the polyphenols identified, in strawberry (cells vs. fruits) and blackberry (cells vs. fruits). Of the total secondary metabolites identified in the samples, at least 26% (not shown) were classified as polyphenols, namely phenolic acids, flavonoids, flavonols, anthocyanins, tannins, lignans and coumarins. For Example, Table 3 shows that out of the polyphenolic compounds identified in the strawberry samples, the majority were identified as phenolic acids (89-82%), and only 0.04 as anthocyanins and 0.04 as tannins. A similar distribution is shown also for the blackberry samples. Table 3- Polyphenolic subclasses distribution in berry cell cultures vs. typical strawberry ripe fruit.

[0163] *Percentages are based on mass spectrometry analysis, and represent the amount of each group out of the total polyphenols identified in the samples.

[0164] Example 6: Taste and Aroma of berry cell culture.

[0165] Under laboratory conditions, 20 strawberry cell culture samples (approximately 3 g fresh weight) were collected from growth vessels and thoroughly rinsed with running tap water to remove residual growth media. A sensory evaluation was then conducted by a panel of 10 judges. All panelists reported that the fresh samples were tasteless, with no indication of sweetness.

[0166] In a follow-up experiment, 13 cell culture samples were dried through lyophilization and presented to the same panel for tasting. Nine out of 10 panelists described the dried samples as not sweet.

[0167] Example 7: Antioxidant activity of berry cell culture

[0168] Testing by ORAC assay

[0169] The acellular Reactive Oxygen Species (ROS) scavenging capacity of a polyphenols extract of strawberry cells, prepared as described above, was evaluated by Oxygen Radical Absorbance Capacity (ORAC) assay, and compared to a polyphenol extract from fresh strawberry fruits. Activity in Trolox equivalents per gram of dried extract was measured in the tested items. In addition, the ORAC values were normalized to amounts of total polyphenols present in the extracts. The normalized results present a significantly higher ORAC activity per amount of polyphenols in the strawberry cell extract compared with the strawberry fruit (Fig. 4). Testing by DC-FDA assay in HaCat cells

[0170] Inhibition of UVB -induced ROS by a polyphenol extract of strawberry cells was tested in HaCaT cells. First, dose-response analysis aimed to determine the highest concentration of each tested item tolerated by the HaCaT cells without causing a reduction in cell viability (i.e. not toxic to the cells). The assay was carried out in triplicates, and each test Item was tested at four concentrations. Polyphenolic extracts were dissolved in DMSO. HaCaT cells (2xl05cells / ml, by counting) were seeded in 96-well plates containing 200 p l / well of complete growth medium. The cells were incubated at 37°C with 5% CO2 until the desired confluency (visual estimation). Then, the medium was aspirated, and the cells were treated w / o or with the test items (4 concentrations), pre-prepared in the growth medium. Naive cells served as a non-treated-basal stage control. 0.1% SDS served as a positive control for a reduction in cell viability (not shown). The vehicle test group served as a negative control. A Blank control group (cells that were not incubated with the MTT solution) was included in the assay. The cells were incubated at 37°C with 5% CO2 for 24h. At the end of the incubation period, the viability of the cells was measured using the MTT assay. Viability greater than 85% was considered nontoxic.

[0171] Extracts of Strawberry fruit and strawberry cells did not affect the cell viability when applied in all the tested concentrations (Fig. 5).

[0172] Next, the ability of the different polyphenolic extracts to inhibit the formation of ROS in HaCaT cells was assessed. Each sample was assayed at four selected concentrations. The assay was carried out in triplicates. HaCaT cells (approx. 2xl05cells / ml, by counting) were seeded in 96 well plates containing 200 pL / well of complete growth medium, and incubated at 37 °C with 5% CO2 until reached 60% confluences (visual estimation). After the cells reached the required confluency, the medium was aspirated, and the cells were mounted with lOOpl DC-FDA (2', 7'- dichlorofluorescein diacetate, 50 pM in PBS) for 30 min. Then, cells were washed once with PBS and treated with lOOpl of preprepared PBS containing various dilutions of the extracts. Blank control groups (cells that were not incubated with the DC-FDA solution) were included in the assay. The following control groups were also added: Naive cells (no UV treatment), stimulated control (irradiation without DMSO), stimulated vehicle control (irradiation and DMSO as a carrier control), and positive control for ROS scavenging (NAC, N-acetylcysteine). The cells were incubated for 15 min at 37°C with 5% CO2, and then exposed to UVB irradiation (20mJ / cm2). Immediately after the exposure to UVB, the generation of ROS was quantified fluorescently (excitation 485nm; emission 535nm; 50 flashes, top reading) and the blank read was subtracted from the fluorescent read. Inhibition of ROS generation was calculated as a percent of the stimulated control. The results are presented in Fig. 6 As can be seen from Fig. 6, strawberry fruit extract at dilutions of 1:8,000 had no visible effect on cellular ROS level and a significant effect was shown only in lower dilutions of 1,1000 (Fig. 6A). The cell culture extract, on the other hand, increased ROS levels at low dilutions (not shown), but inhibited ROS formation at higher dilutions (Fig. 6B). A dilution of about 1:100,000 of the cell culture extract provided ROS formation inhibition of about 16%, which was similar to the results obtained using 1:1,000 dilution of fruit extract. This indicates that the cell culture extract, at the right dilution, is much more potent in ROS inhibition than fruit extract.

[0173] Relationship between antioxidant capacity and extract polyphenol content

[0174] To better understand the contribution of the antioxidant efficacy compared to polyphenols present in the extracts, significant ORAC results obtained by all extracts were normalized to the amount of total polyphenols in the samples. It was found that while the strawberry fruit extract contained 1.9% polyphenols, the strawberry cell culture extract contained 0.8% polyphenols. Additionally, the % polyphenols in the diluted extract that was needed to obtain 15% inhibition of UVB-induced ROS in the DC-FDA assay was calculated and found to be about 2.6 X 10’3for the fruit extract, and about 7.6 X 10’6for the cell culture extract. That is, polyphenols extracted from the strawberry cell culture were more potent compared to fruit extract, as a significantly lower amount of polyphenols was needed to inhibit ROS formation.

Claims

CLAIMS1. A composition comprising culture-derived berry cells, wherein the culture-derived berry cells contain a polyphenolic fraction comprising at least about 50% phenolic acids.

2. The composition of claim 1, wherein the culture-derived berry cells comprise at least about 12% w / w protein of dry cells weight.

3. The composition of claim 1 or 2, wherein the culture-derived berry cells comprise less than about 20% w / w sugar of dry cells weight.

4. The composition of any one of claims 1-3, wherein the culture-derived berry cells comprise at least about 0.5% w / w polyphenols of dry cells weight.

5. The composition of any one of claims 1-4, wherein the culture-derived berry cells comprise at least about 20% w / w dietary fibers of dry cells weight.

6. The composition of any one of claims 1-5, wherein the culture-derived berry cells comprise a secondary metabolites fraction comprising at least about 30% polyphenols.

7. The composition of any one of claims 1-6, wherein the culture-derived berry cells comprise a secondary metabolites fraction comprising at least about 25% phenolic acids.

8. The composition of any one of claims 1-7, wherein the culture-derived berry cells comprise a secondary metabolites fraction comprising of at least about 20% terpenoids.

9. The composition of any one of claims 1-8, wherein the culture-derived berry cells comprise a secondary metabolites fraction comprising of at least about 10% alkaloids.

10. The composition of any one of claims 1-9, wherein the polyphenolic fraction of the culture- derived berry cells comprises at least about 60% non-flavonoids polyphenols.

11. The composition of any one of claims 1-10, wherein the polyphenolic fraction of the culture- derived berry cells comprises at least about 80% phenolic acids.

12. The composition of any one of claims 1-11, wherein the polyphenolic fraction of the culture- derived berry cells comprises less than about 10% flavonoids.

13. The composition of any one of claims 1-12, wherein the polyphenolic fraction of the culture- derived berry cells comprises less than about 5% anthocyanins.

14. The composition of any one of claims 1-13, wherein the polyphenolic fraction of the culture- derived berry cells comprises less than about 5% tannins.

15. The composition of any one of claims 1-14, wherein the polyphenolic fraction of the culture- derived berry cells comprises at least about 1% lignans and coumarins.

16. The composition of any one of claims 1-15, wherein the composition is in the form of a dry powder.

17. The composition of claim 16, wherein the dry powder is a freeze-dried, a spray dried, or a fluid bed dried powder.

18. The composition of any one of claims 1-15, wherein the composition is in the form of a liquid suspension.

19. The composition of any one of claims 1-18, comprising at least about 70% w / w whole berry cells.

20. The composition of one of claims 1-19, wherein the composition is edible.

21. The composition of one of claims 1-20, wherein the composition is not sweet.

22. The composition of any one of claims 1-21, wherein the berry cells are selected from strawberry cells, blackberry cells, blueberry cells, chokeberry cells, elderberry cells, raspberry cells, and a combination thereof.

23. The composition of any one of claims 1- 22, wherein the berry cells are from a genus selected from Fragaria, Rubus, Vaccinium, Aronia, Sambucus, and a combination thereof.

24. The composition of claim 23, wherein the berry cells are from Fragaria x ananassa and / or Rubus Fruticosus.

25. The composition of any one of claims 1-24, further comprising a cell culture medium.

26. The composition of claim 25, wherein the medium comprises a hormone, 2- Morpholinoethanesulfonic acid monohydrate (MES), and / or amino acids.

27. A food product comprising the composition of any one of claims 1-24.

28. A food supplement comprising the composition of any one of claims 1-24.

29. A nutraceutical comprising the composition of any one of claims 1-24.

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