Expansion of bovine skeletal muscle stem cells from spinner flasks to benchtop stirred-tank bioreactors for up to 38 days.

Introduction: Successful long-term expansion of skeletal muscle satellite cells (MuSCs) on a large scale is fundamental for cultivating animal cells for protein production. Prerequisites for efficient cell expansion include maintaining essential native cell activities such as cell adhesion, migration, proliferation, and differentiation while ensuring consistent reproducibility. Method: This study investigated the growth of bovine MuSC culture using low-volume spinner flasks and a benchtop stirred-tank bioreactor (STR). Results and discussion: Our results showed for the first time the expansion of primary MuSCs for 38 days in a bench-top STR run with low initial seeding density and FBS reduction, supported by increased expression of the satellite cell marker PAX7 and reduced expression of differentiation-inducing genes like MYOG, even without adding p38-MAPK inhibitors. Moreover, the cells retained their ability to proliferate, migrate, and differentiate after enzymatic dissociation from the microcarriers. We also showed reproducible results in a separate biological benchtop STR run.

Production of food-grade microcarriers based on by-products from the food industry to facilitate the expansion of bovine skeletal muscle satellite cells for cultured meat production.

A major challenge for successful cultured meat production is the requirement for large quantities of skeletal muscle satellite cells (MuSCs). Commercial microcarriers (MCs), such as Cytodex®1, enable extensive cell expansion by offering a large surface-to-volume ratio. However, the cell-dissociation step post cell expansion makes the cell expansion less efficient. A solution is using food-grade MCs made of sustainable raw materials that do not require a dissociation step and can be included in the final meat product. This study aimed to produce food-grade MCs from food industry by-products (i.e., turkey collagen and eggshell membrane) and testing their ability to expand bovine MuSCs in spinner flask systems for eight days. The MCs' physical properties were characterized, followed by analyzing the cell adhesion, growth, and metabolic activity. All MCs had an interconnected porous structure. Hybrid MCs composed of eggshell membrane and collagen increased the mechanical hardness and stabilized the buoyancy compared to pure collagen MCs. The MuSCs successively attached and covered the entire surface of all MCs while expressing high cell proliferation, metabolic activity, and low cell cytotoxicity. Cytodex®1 MCs were included in the study. Relative gene expression of skeletal muscle markers showed reduced PAX7 and increased MYF5, which together with augmented proliferation marker MKI67 indicated activated and proliferating MuSCs on all MCs. Furthermore, the expression pattern of cell adhesion receptors (ITGb5 and SDC4) and focal adhesion marker VCL varied between the distinct MCs, indicating different specific cell receptor interactions with the various biomaterials. Altogether, our results demonstrate that these biomaterials are promising prospects to produce custom-fabricated food-grade MCs intended to expand MuSCs.

Screening of by-products from the food industry as growth promoting agents in serum-free media for skeletal muscle cell culture.

The most significant cost driver for efficient bio-production of edible animal proteins is the cell culture media, where growth factors account for up to 96% of the total cost. The culture media must be serum-free, affordable, contain only food-grade ingredients, be efficient to promote cell growth and available in massive quantities. The commercially available serum substitutes are expensive and not necessarily food-grade. Identifying inexpensive food-safe alternatives to serum is crucial. By-products from food production are available in massive quantities, contain potential factors that can promote growth and are promising ingredients for serum replacement. The main goal of this study was to explore if food-grade by-product materials can be used as growth promoting agents in skeletal muscle cell culture to develop a tailor-made serum free media. Different by-products, including chicken carcass, cod backbone, eggshell membrane, egg white powder and pork plasma were enzymatically or chemically hydrolyzed. The hydrolysates in addition to lyophilized pork plasma and yeast extract were further characterized by size-exclusion chromatography, elemental combustion analysis and degree of hydrolysis. The materials were used as supplement to or replacement of commercial serum and further evaluated for their effect on metabolic activity, cell proliferation and cell cytotoxicity in muscle cells cultured in vitro. Our results indicate that none of the materials were cytotoxic to the skeletal muscle cells. Hydrolysates rich in peptides with approximately 2-15 amino acids in length were shown to improve cell growth and metabolic activity. Of all the materials tested pork plasma hydrolysates and yeast extract were the most promising. Pork plasma hydrolysates increased metabolic activity by 110% and cell proliferation with 48% when cultured in serum-free conditions for 3 days compared with control cells cultured with full serum conditions. Most interestingly, this response was dependent on both material and choice of enzyme used. We suggest that these materials have the potential to replace serum during cultivation and as such be included in a tailor-made serum-free media.

First Observation of CP Violation in B[over ¯]

We report a measurement of the time-dependent CP asymmetry of B[over ¯]^{0}→D_{CP}^{(*)}h^{0} decays, where the light neutral hadron h^{0} is a π^{0}, η, or ω meson, and the neutral D meson is reconstructed in the CP eigenstates K^{+}K^{-}, K_{S}^{0}π^{0}, or K_{S}^{0}ω. The measurement is performed combining the final data samples collected at the ϒ(4S) resonance by the BABAR and Belle experiments at the asymmetric-energy B factories PEP-II at SLAC and KEKB at KEK, respectively. The data samples contain (471±3)×10^{6} BB[over ¯] pairs recorded by the BABAR detector and (772±11)×10^{6} BB[over ¯] pairs recorded by the Belle detector. We measure the CP asymmetry parameters -η_{f}S=+0.66±0.10(stat)±0.06(syst) and C=-0.02±0.07(stat)±0.03(syst). These results correspond to the first observation of CP violation in B[over ¯]^{0}→D_{CP}^{(*)}h^{0} decays. The hypothesis of no mixing-induced CP violation is excluded in these decays at the level of 5.4 standard deviations.

Observation of the B(s)(0)→η'η' Decay.

The first observation of the B(s)(0)→η'η' decay is reported. The study is based on a sample of proton-proton collisions corresponding to 3.0 fb(-1) of integrated luminosity collected with the LHCb detector. The significance of the signal is 6.4 standard deviations. The branching fraction is measured to be [3.31±0.64(stat)±0.28(syst)±0.12(norm)]×10(-5), where the third uncertainty comes from the B(±)→η'K(±) branching fraction that is used as a normalization. In addition, the charge asymmetries of B(±)→η'K(±) and B(±)→ϕK(±), which are control channels, are measured to be (-0.2±1.3)% and (+1.7±1.3)%, respectively. All results are consistent with theoretical expectations.

Measurement of the [Formula: see text] production cross-section in proton-proton collisions via the decay [Formula: see text].

The production of the [Formula: see text] state in proton-proton collisions is probed via its decay to the [Formula: see text] final state with the LHCb detector, in the rapidity range [Formula: see text] and in the meson transverse-momentum range [Formula: see text]. The cross-section for prompt production of [Formula: see text] mesons relative to the prompt [Formula: see text] cross-section is measured, for the first time, to be [Formula: see text] at a centre-of-mass energy [Formula: see text] using data corresponding to an integrated luminosity of 0.7 fb[Formula: see text], and [Formula: see text] at [Formula: see text] using 2.0 fb[Formula: see text]. The uncertainties quoted are, in order, statistical, systematic, and that on the ratio of branching fractions of the [Formula: see text] and [Formula: see text] decays to the [Formula: see text] final state. In addition, the inclusive branching fraction of [Formula: see text]-hadron decays into [Formula: see text] mesons is measured, for the first time, to be [Formula: see text], where the third uncertainty includes also the uncertainty on the [Formula: see text] inclusive branching fraction from [Formula: see text]-hadron decays. The difference between the [Formula: see text] and [Formula: see text] meson masses is determined to be [Formula: see text].

Search for long-lived particles decaying to jet pairs.

A search is presented for long-lived particles with a mass between 25 and 50 [Formula: see text] and a lifetime between 1 and 200[Formula: see text] in a sample of proton-proton collisions at a centre-of-mass energy of [Formula: see text] TeV, corresponding to an integrated luminosity of 0.62 [Formula: see text], collected by the LHCb detector. The particles are assumed to be pair-produced by the decay of a standard model-like Higgs boson. The experimental signature of the long-lived particle is a displaced vertex with two associated jets. No excess above the background is observed and limits are set on the production cross-section as a function of the long-lived particle mass and lifetime.

Measurement of Bc+ Production in Proton-Proton Collisions at √[s]=8 TeV.

Production of Bc+ mesons in proton-proton collisions at a center-of-mass energy of 8 TeV is studied with data corresponding to an integrated luminosity of 2.0 fb-1 recorded by the LHCb experiment. The ratio of production cross sections times branching fractions between the Bc+→J/ψπ+ and B+→J/ψK+ decays is measured as a function of transverse momentum and rapidity in the regions 0

Observation of two new Ξ(b)(-) baryon resonances.

Two structures are observed close to the kinematic threshold in the Ξ(b)(0)π(-) mass spectrum in a sample of proton-proton collision data, corresponding to an integrated luminosity of 3.0 fb(-1), recorded by the LHCb experiment. In the quark model, two baryonic resonances with quark content bds are expected in this mass region: the spin-parity J(P)=(1/2)(+) and J(P)=(3/2)(+) states, denoted Ξ(b)('-) and Ξ(b)(*-). Interpreting the structures as these resonances, we measure the mass differences and the width of the heavier state to be m(Ξ(b)('-))-m(Ξ(b)(0))-m(π(-))=3.653±0.018±0.006 MeV/c(2), m(Ξ(b)(*-))-m(Ξ(b)(0))-m(π(-))=23.96±0.12±0.06 MeV/c(2), Γ(Ξ(b)(*-))=1.65±0.31±0.10 MeV, where the first and second uncertainties are statistical and systematic, respectively. The width of the lighter state is consistent with zero, and we place an upper limit of Γ(Ξ(b)('-))<0.08 MeV at 95% confidence level. Relative production rates of these states are also reported.

Measurement of the semileptonic CP asymmetry in B0-B[over ¯]0 mixing.

The semileptonic CP asymmetry in B0-B[over ¯]0 mixing, a(sl)(d), is measured in proton-proton collision data, corresponding to an integrated luminosity of 3.0 fb-1, recorded by the LHCb experiment. Semileptonic B0 decays are reconstructed in the inclusive final states D-µ+ and D*-µ+, where the D- meson decays into the K+π-π- final state and the D*- meson into the D[over ¯]0(→K+π-)π- final state. The asymmetry between the numbers of D(*)-µ+ and D(*)+µ- decays is measured as a function of the decay time of the B0 mesons. The CP asymmetry is measured to be a(sl)(d)=(-0.02±0.19±0.30)%, where the first uncertainty is statistical and the second systematic. This is the most precise measurement of a(sl)(d) to date and is consistent with the prediction from the standard model.

Precision measurement of CP violation in B(S)(0)→J/ΨK+K- decays.

The time-dependent CP asymmetry in B(s)(0)→J/ψK+K- decays is measured using pp collision data, corresponding to an integrated luminosity of 3.0 fb-1, collected with the LHCb detector at center-of-mass energies of 7 and 8 TeV. In a sample of 96,000 B(s)(0)→J/ψK+K- decays, the CP-violating phase ϕs is measured, as well as the decay widths ΓL and ΓH of the light and heavy mass eigenstates of the B(s)(0)-B[over ¯]s0 system. The values obtained are ϕs=-0.058±0.049±0.006 rad, Γs≡(ΓL+ΓH)/2=0.6603±0.0027±0.0015 ps-1, and ΔΓs≡ΓL-ΓH=0.0805±0.0091±0.0032 ps-1, where the first uncertainty is statistical and the second, systematic. These are the most precise single measurements of those quantities to date. A combined analysis with Bs0→J/ψπ+π- decays gives ϕs=-0.010±0.039 rad. All measurements are in agreement with the standard model predictions. For the first time, the phase ϕs is measured independently for each polarization state of the K+K- system and shows no evidence for polarization dependence.

Precision measurement of the mass and lifetime of the Ξb⁻ baryon.

We report on measurements of the mass and lifetime of the Ξ(b)⁻ baryon using about 1800 Ξ(b)⁻ decays reconstructed in a proton-proton collision data set corresponding to an integrated luminosity of 3.0 fb⁻¹ collected by the LHCb experiment. The decays are reconstructed in the Ξ(b)⁻→Ξ(c)°π⁻, Ξ(c)°â†’pK⁻K⁻π⁺ channel and the mass and lifetime are measured using the Λ(b)°â†’Λ(c)⁺π⁻ mode as a reference. We measure M(Ξ(b)⁻)-M(Λ(b)°)=178.36±0.46±0.16 MeV/c², (τ(Ξ(b)⁻)/τ(Λ(b)°)=1.089±0.026±0.011, where the uncertainties are statistical and systematic, respectively. These results lead to a factor of 2 better precision on the Ξ(b)⁻ mass and lifetime compared to previous best measurements, and are consistent with theoretical expectations.

Measurement of the CP-violating phase ϕs in Bs(0)→Ds(+)Ds(-) decays.

We present a measurement of the CP-violating weak mixing phase ϕs using the decay Bs(0)→Ds(+)Ds(-) in a data sample corresponding to 3.0 fb(-1) of integrated luminosity collected with the LHCb detector in pp collisions at center-of-mass energies of 7 and 8 TeV. An analysis of the time evolution of the system, which does not use the constraint |λ|=1 to allow for the presence of CP violation in decay, yields ϕs=0.02±0.17(stat)±0.02(syst) rad, |λ|=0.91(-0.15)(+0.18)(stat)±0.02(syst). This result is consistent with the standard model expectation.

Measurement of the B¯s° meson lifetime in Ds⁺π⁻ decays.

We present a measurement of the ratio of the B¯s° meson lifetime, in the flavor-specific decay to Ds⁺π⁻, to that of the B¯° meson. The pp collision data used correspond to an integrated luminosity of 1 fb(-1), collected with the LHCb detector, at a center-of-mass energy of 7 TeV. Combining our measured value of 1.010±0.010±0.008 for this ratio with the known B¯° lifetime, we determine the flavor-specific B¯s° lifetime to be τ(B¯s° )=1.535±0.015±0.014 ps, where the uncertainties are statistical and systematic, respectively. This is the most precise measurement to date, and is consistent with previous measurements and theoretical predictions.

Test of lepton universality using b+ → K+ℓ+ℓ- decays.

A measurement of the ratio of the branching fractions of the B(+) → K(+)µ(+)µ(-) and B(+) → K(+)e(+)e(-) decays is presented using proton-proton collision data, corresponding to an integrated luminosity of 3.0 fb(-1), recorded with the LHCb experiment at center-of-mass energies of 7 and 8 TeV. The value of the ratio of branching fractions for the dilepton invariant mass squared range 1 < q(2) < 6 GeV(2)/c(4) is measured to be 0.745(-0.074)(+0.090)(stat) ± 0.036(syst). This value is the most precise measurement of the ratio of branching fractions to date and is compatible with the standard model prediction within 2.6 standard deviations.

First observation of a baryonic Bc+ decay.

A baryonic decay of the B(c)(+) meson, B(c)(+) → J/ψppπ(+), is observed for the first time, with a significance of 7.3 standard deviations, in pp collision data collected with the LHCb detector and corresponding to an integrated luminosity of 3.0 fb(-1) taken at center-of-mass energies of 7 and 8 TeV. With the B(c)(+) → J/ψπ(+) decay as the normalization channel, the ratio of branching fractions is measured to be B(B(c)(+) → J/ψppπ(+))/B(B(c)(+) → J/ψπ(+)) = 0.143(-0.034)(+0.039)(stat) ± 0.013(syst). The mass of the B(c)(+) meson is determined as M(B(c)(+) = 6274.0 ± 1.8(stat) ± 0.4(syst) MeV/c(2), using the B(c)(+) → J/ψppπ(+) channel.

Observation of overlapping spin-1 and spin-3 D0K- resonances at mass 2.86 GeV/c2.

The resonant substructure of B(s)(0) → D(0)K(-)π(+) decays is studied using a data sample corresponding to an integrated luminosity of 3.0 fb(-1) of pp collision data recorded by the LHCb detector. An excess at m(D(0)K(-))≈ 2.86 GeV/c(2) is found to be an admixture of spin-1 and spin-3 resonances. Therefore, the D(sJ)*(2860)(-) state previously observed in inclusive e(+)e(-) → D(0)K(-)X and pp → D(0)K(-)X processes consists of at least two particles. This is the first observation of a heavy flavored spin-3 resonance, and the first time that any spin-3 particle has been seen to be produced in B decays. The masses and widths of the new states and of the D(s2)*(2573)(-) meson are measured, giving the most precise determinations to date.

Evidence for CP violation in B+ → ppK+ decays.

Three-body B+ → ppK+ and B(+) → ppπ(+) decays are studied using a data sample corresponding to an integrated luminosity of 3.0 fb(-1) collected by the LHCb experiment in proton-proton collisions at center-of-mass energies of 7 and 8 TeV. Evidence of CP violation in the B(+) → ppK(+) decay is found in regions of the phase space, representing the first measurement of this kind for a final state containing baryons. Measurements of the forward-backward asymmetry of the light meson in the pp rest frame yield A(FB)(ppK(+),m(pp)<2.85 GeV/c(2)) = 0.495 ± 0.012 (stat) ± 0.007 (syst) and A(FB)(ppπ(+),m(pp) <2.85 GeV/c(2)) = -0.409 ± 0.033 (stat) ± 0.006 (syst). In addition, the branching fraction of the decay B(+) → Λ(1520)p is measured to be B(B(+) → Λ(1520)p) = (3.15 ± 0.48 (stat) ± 0.07 (syst) ± 0.26 (BF)) × 10(-7), where BF denotes the uncertainty on secondary branching fractions.

First measurement of the charge asymmetry in beauty-quark pair production.

The difference in the angular distributions between beauty quarks and antiquarks, referred to as the charge asymmetry, is measured for the first time in bb pair production at a hadron collider. The data used correspond to an integrated luminosity of 1.0 fb(-1) collected at 7 TeV center-of-mass energy in proton-proton collisions with the LHCb detector. The measurement is performed in three regions of the invariant mass of the bb system. The results obtained are A(C)(bb))(40 105 GeV/c(2)) = 1.6 ± 1.7 ± 0.6%, where A(C)(bb)) is defined as the asymmetry in the difference in rapidity between jets formed from the beauty quark and antiquark, where in each case the first uncertainty is statistical and the second systematic. The beauty jets are required to satisfy 2 < η < 4, E(T) >20 GeV, and have an opening angle in the transverse plane Δ Ï• > 2.6 rad. These measurements are consistent with the predictions of the standard model.

Precision measurement of the mass and lifetime of the Ξ(b)(0) baryon.

Using a proton-proton collision data sample corresponding to an integrated luminosity of 3 fb(-1) collected by LHCb at center-of-mass energies of 7 and 8 TeV, about 3800 Ξ(b)(0) → Ξ(c)(+)π(-), Ξ(c)(+)) → pK(-)π(+) signal decays are reconstructed. From this sample, the first measurement of the Ξ(b)(0) baryon lifetime is made, relative to that of the Λ(b)(0) baryon. The mass differences M(Ξ(b)(0))-M(Λ(b)(0)) and M(Ξ(c)(+))-M(Λ(c)(+)) are also measured with precision more than 4 times better than the current world averages. The resulting values are τ(Ξ(b)(0))/τ(Λ)(b)(0)) = 1.006 ± 0.018 ± 0.010,M(Ξ(b)(0))-M(Λ(b)(0)) = 172.44 ± 0.39 ± 0.17 MeV/c(2),M(Ξ(c)(+))-M(Λ(c)(+)) = 181.51 ± 0.14 ± 0.10 MeV/c(2),where the first uncertainty is statistical and the second is systematic. The relative rate of Ξ(b)(0) to Λ(b)(0) baryon production is measured to be f(Ξ)(b)(0))/f(Λ)(b)(0))B(Ξ(b)(0) → Ξ(c)(+)π(-))/B(Λ(b)(0) → Λ(c)(+)π(-))B(Ξ(c)(+) → pK(-)π(+))/B(Λ(c)(+) → pK(-)}π(+)) = (1.88 ± 0.04 ± 0.03) × 10(-2),where the first factor is the ratio of fragmentation fractions, b → Ξ(b)(0) relative to b → Λ(b)(0). Relative production rates as functions of transverse momentum and pseudorapidity are also presented.