Optimizing chicken muscle stem cell isolation using recombinant proteolytic enzymes for cultured meat production
Jeong Ho Lim, Sibhghatulla Shaikh, Woo-Jong Lee, Eun Ju Lee, Inho Choi
TL;DR
This paper shows that using recombinant enzymes improves the isolation of chicken muscle stem cells, which is important for cultured meat production.
Contribution
The study introduces recombinant proteolytic enzymes as a safer and more efficient method for isolating muscle stem cells compared to traditional crude enzymes.
Findings
Recombinant enzymes significantly improved tissue dissociation and increased the yield of muscle stem cells.
MSCs isolated with recombinant enzymes showed better proliferation and differentiation characteristics.
The method offers a safer and more efficient alternative for stem cell isolation in cultured meat production.
Abstract
Muscle satellite cells, also known as muscle stem cells (MSCs), are crucial for muscle growth and regeneration, and are among the most commonly used cell types for cultured meat production. Efficient isolation of high-quality MSCs is essential for both basic research and for the development of therapies targeting muscle-related diseases and for advancing cultured meat production. Traditional isolation methods often use crude proteolytic enzymes, which can lead to contamination and cytotoxicity. This study aimed to improve the isolation of muscle stem cells by using recombinant proteolytic enzymes—collagenase and thermolysin—while comparing their performance to crude enzyme treatments. The isolation of MSCs from chicken muscle tissue was conducted using both crude pronase and recombinant enzymes. The effectiveness of each method was evaluated based on tissue dissociation efficiency and…
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Taxonomy
TopicsMuscle Physiology and Disorders · Meat and Animal Product Quality · Animal Genetics and Reproduction
Introduction
Muscle tissue is essential for various physiological functions, including movement, posture, and involuntary actions. It is classified into three primary types: skeletal, cardiac, and smooth muscle, each with distinct structural and functional characteristics. Skeletal muscle, the most abundant type, accounts for approximately 30–40% of total body weight. This striated, voluntary muscle is attached to bones via tendons and plays a key role in force generation and movement through contraction (Allen et al. 2024).
Muscle satellite cells, also known as muscle stem cells (MSCs), are vital for the growth, repair, and regeneration of skeletal muscles. First identified by Mauro in 1961, these cells reside between the basal lamina and sarcolemma of muscle fibers (Mauro 1961). MSCs typically remain in a quiescent state but become activated in response to muscle injury or physiological stress, such as exercise. Upon activation, they proliferate and differentiate into myoblasts, which then fuse to form new muscle fibers or repair damaged ones (Li et al. 2025). Regeneration of skeletal muscle relies heavily on MSCs, which regulate myofiber development through key myogenic transcription factors such as paired box protein 3 (Pax3), paired box protein 7 (Pax7), myogenic differentiation 1 (MYOD), and myogenin (MYOG) (Pang et al. 2023). Upon injury, Pax7^+^ MSCs are activated, inducing the expression of MYOD, myogenic factor 5 (Myf5), and MYOG, which drive myotube formation (Yu et al. 2021). Various extracellular matrix (ECM) components, such as fibromodulin (Ahmad et al. 2023; Lee et al. 2021a), matrix gla protein (Ahmad et al. 2017), and dermatopontin (Kim et al. 2019), as well as cell adhesion molecules like immunoglobulin-like cell adhesion molecules 4 and 5 (Lim et al. 2021, 2022), also regulate myogenesis and facilitate skeletal muscle regeneration.
Cultured meat has emerged as a promising alternative to traditional livestock farming, addressing major concerns such as poor breeding conditions, environmental pollution from wastewater and methane emissions, and animal welfare issues (Lee et al. 2023). Cultured meat production involves a stepwise process that begins with the isolation of MSCs from target livestock muscle tissue using enzymatic digestion. Isolated MSCs undergo proliferation and subsequent myogenic differentiation, eventually leading to the formation of cultured meat (Lee et al. 2021b). Obtaining high-quality MSCs at the initial stage is critical because it has a significant impact on the efficiency of cell expansion, differentiation, and the overall quality of the final cultured meat product.
The study of MSCs is crucial for elucidating the mechanisms of muscle development and for improving the efficiency of cultured meat production. MSCs were first isolated from adult rat skeletal muscle by Bischoff in 1974 (Bischoff 1974). Enzymatic treatments such as pronase and trypsin were used to extract viable MSCs from muscle tissue. This approach, with a few modifications, is still used to isolate avian MSCs for cell culture (Velleman 2024). Isolating high-quality MSCs with high viability, purity, and myogenic potential from muscle tissue is critical for advancing these studies. The most common isolation method involves finely mincing muscle tissue and extracting MSCs using proteolytic enzymes such as collagenase, dispase, and pronase (Motohashi et al. 2014). Collagenase, derived from Clostridium hystolyticum, is the most widely used enzyme in this process. In addition, collagenase plays a pivotal role in tissue engineering and clinical applications due to its ability to efficiently dissociate tissues and extract cells. However, concerns remain regarding the safety and stability of cells isolated using crude collagenase, primarily due to its pathogenic origin (Xu et al. 2025). Even crude type collagenase is a complex mixture of collagen-degrading enzymes, neutral proteases, and other enzymatic components. However, the variability in composition and activity of these crude formulations can affect the consistency and efficiency of cell isolation. Therefore, the use of highly purified recombinant collagenase is increasingly recommended to ensure reproducibility, safety, and high-quality outcomes (Fujio et al. 2014). Recombinant collagenase, produced through recombinant DNA technology in Escherichia coli, offers a safer alternative to the crude form. Recombinant collagenase, particularly Collagenase Class I (Col G) and Class II (Col H), has been shown to provide more consistent and sustainable results in various cell isolation protocols, including those for cardiomyocytes (Campora et al. 2018).
Thermolysin, another protease of interest, is a thermostable neutral metalloprotease derived from Bacillus thermoproteolyticus. It selectively cleaves peptide bonds near hydrophobic amino acids and efficiently degrades ECM components such as collagen, elastin, and fibronectin while preserving cell surface proteins. Its low nonspecific activity, thermostability, and ECM-degrading capacity make it a suitable complement to collagenase in cell isolation protocols (Perreault and Beaulieu 1996).
In this study, recombinant collagenase was used to target collagen, the principal component of muscle connective tissue and fascia, in combination with thermolysin, which degrades a broader range of ECM components. This enzymatic approach was applied to isolate MSCs from chicken muscle tissue for improved yield and quality in cultured meat production.
Materials and methods
Animal experiment
All experiments were approved by the Institutional Animal Care and Use Committee of Yeungnam University (Approval No. AEC2022-022) and conducted in accordance with its guidelines.
Isolation and culture of chicken MSCs
Whole leg muscles were harvested from 16-d-old embryos, minced, and enzymatically digested for 1 h at 37 °C using 0.1% Pronase (Roche, Mannheim, Germany), 0.1% TDzyme C, 0.06% TDzyme T, or a combination of 0.015% TDzyme C and 0.03% TDzyme T (TDzyme CT) (CONNEXT, Daegu, Korea). The digested tissue was centrifuged at 1,000 × g for 3 min, filtered through a 40-µm cell strainer (Corning, Brooklyn, NY, USA), and centrifuged again at 1,000 × g for 5 min. The resulting pellet was resuspended in Ham’s F-10 medium supplemented with 20% fetal bovine serum (FBS; Cytiva, Marlborough, MA, USA), 1% penicillin–streptomycin (P/S; Cytiva, Marlborough, MA, USA), and 5 ng/mL fibroblast growth factor 2 (FGF2; Miltenyi Biotec, Bergisch Gladbach, Germany). Cells were seeded onto collagen-coated plates (Corning, Brooklyn, NY, USA) and cultured at 37 °C in an incubator with 5% CO_2_. When the cells reached over 90% confluency on the surface of the culture dish, the medium was replaced with DMEM containing 1% penicillin–streptomycin and 2% FBS to induce differentiation.
Cell viability analysis based on acridine orange and propidium iodide (AO/PI) staining
Cell viability was determined by staining with acridine orange and propidium iodide (AO/PI) (Thermo Fisher Scientific, Waltham, MA, USA) to distinguish between living and dead cells. Following staining, the samples were analyzed on the Countess 3 FL Automated Cell Counter (Thermo Fisher Scientific, Waltham, MA, USA), which uses fluorescence detection to accurately quantify viable and non-viable cells based on membrane integrity. All procedures were performed according to the manufacturer’s instructions.
Methyltetrazolium salt (MTS)-based cell proliferation assay
Cell proliferation was assessed using the CellTiter 96 AQueous One Solution Reagent (Promega, Madison, WI, USA). Briefly, cells were incubated with the reagent for 1 h at 37 °C in an incubator with 5% CO_2_. Absorbance was measured at 490 nm using a microplate reader (Biotek Synergy H1, Winooski, VT, USA).
Creatine kinase activity assay
To assess myogenic differentiation, creatine kinase (CK) activity was measured (Lee et al. 2025) using the EnzyChrom Creatine Kinase Assay Kit (BioAssay Systems, Hayward, CA, USA), following the manufacturer’s instructions.
Immunocytochemistry
MSCs were cultured in glass-bottom dishes with differentiation media for 4 d. Subsequently, the cells were rinsed twice with phosphate-buffered saline (PBS) and fixed with 4% formaldehyde for 10 min. The cells were then permeabilized using 0.2% Triton X-100 (Sigma Aldrich, St. Louis, MO, USA) and incubated overnight with anti-myosin heavy chain (MYH) primary antibody (1:50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4 °C in a humidified environment. Subsequently, the cells were incubated with Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (1:100; Invitrogen, Carlsbad, CA, USA) in the dark for 1 h at room temperature. Following two PBS washes, the nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; 1:1,000; Sigma Aldrich, St. Louis, MO, USA), and fluorescent images were captured using a Nikon Eclipse TE2000-U inverted fluorescence phase contrast microscope (Nikon, Melville, NY, USA).
Real-time reverse transcription polymerase chain reaction (RT-PCR) analysis
Total RNA was extracted using Trizol Reagent (Invitrogen, Carlsbad, CA, USA), and complementary DNA (cDNA) was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA); both procedures were performed according to the respective manufacturers’ instructions. Real-time RT-PCR was performed using Power SYBR Green PCR Master Mix (Applied Biosystems, Foster City, CA, USA) as previously described (Ahmad et al. 2024). The primers used in this study are listed in Supplementary Table S1.
Western blot analysis
MSCs were lysed with RIPA buffer containing 1% protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA). Protein concentrations were quantified using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts (50 μg) of protein were resolved on 8% or 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Burlington, MA, USA). The membranes were blocked with bovine serum albumin (BSA) or 3% skim milk in Tris-buffered saline (TBS) containing 0.1% Tween 20. Subsequently, they were incubated overnight at 4 °C with primary antibodies against MYOD (1:400; Santa Cruz Biotechnology, Dallas, TX, USA), MYOG (1:400; Abcam, Cambridge, UK), MYH (1:400; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or β-actin (1:400; Santa Cruz Biotechnology, Dallas, TX, USA) in TBS with 1% skim milk or BSA. Following washing, the membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (goat anti-mouse or anti-rabbit; Santa Cruz Biotechnology, Santa Cruz Biotechnology, Dallas, TX, USA) for 2 h at room temperature. Bands were visualized using the Dyne ECL Pico Plus Western Blotting Detection Kit (Dyne Bio, Seongnam, South Korea) and imaged using an Azure 300 chemiluminescent imager (Azure Biosystems, Dublin, CA, USA).
Statistical analysis
All data were derived from at least three independent experiments. Statistical comparisons of means were performed using one-way ANOVA with Tukey’s post hoc test via the PROC GLM procedure in SAS version 9.0. Results were considered statistically significant at p < 0.05.
Results
MSC isolation using crude and recombinant proteolytic enzymes
Both crude and recombinant proteolytic enzymes were evaluated for their effectiveness in isolating chicken MSCs. The enzymes were applied to chicken muscle tissue and incubated with agitation at 37 °C for 60 min. The resulting cell suspensions were then passed through a 100-µm cell strainer to separate dissociated cells from undigested tissue. Among the tested enzymes, the crude enzyme Pronase and the recombinant enzyme combination TDzyme CT retained minimal tissue in the strainer, indicating efficient digestion of muscle tissue. In contrast, the recombinant enzymes TDzyme C and TDzyme T, when used individually, retained a substantial amount of undigested tissue, suggesting a lower tissue dissociation capacity (Fig. 1A). The number of MSCs isolated with each enzyme was quantified and expressed as the number of MSCs per gram of muscle tissue. TDzyme CT yielded the highest MSC count, significantly surpassing that obtained with the commonly used Pronase. In comparison, both TDzyme C and TDzyme T resulted in significantly lower MSC yields than Pronase (Fig. 1B). Cell viability was assessed using acridine orange and propidium iodide (AO/PI) staining. All enzyme-treated groups demonstrated high cell viability, with over 95% of cells remaining viable after isolation (Fig. 1C).Fig. 1. Isolation of MSCs using crude and recombinant proteolytic enzymes. (A) Representative images of residual tissue fragments following enzymatic dissociation of chicken muscle and filtration through a 40-µm cell strainer. (B) Quantification of isolated MSCs, expressed as the number of cells per gram of muscle tissue. (C) Cell viability assessment using AO/PI staining
Purity of MSCs using crude or recombinant proteolytic enzymes
To assess the purity of isolated MSCs, expression of the MSC marker PAX7 was evaluated using immunocytochemistry (red: PAX7, blue: nuclei) (Fig. 2A). MSCs isolated using crude (Pronase), recombinant (TDzyme C, TDzyme T), and combined recombinant (TDzyme CT) proteolytic enzymes were all found to have high purity, with PAX7-positive cells exceeding 90% in most groups. The exception was TDzyme C, which yielded a slightly lower proportion of PAX7-positive cells. Among all groups, Pronase and TDzyme CT achieved the highest purity levels, with no significant difference between them. These results confirm that the MSC isolation purity achieved with the commonly used crude enzyme Pronase is comparable to that of the optimized recombinant combination TDzyme CT (Fig. 2B).Fig. 2. Purity of MSCs isolated using crude and recombinant proteolytic enzymes. (A) Immunocytochemical detection of PAX7 (red), a marker of muscle satellite cells; nuclei were counterstained with DAPI (blue). (B) Quantification of PAX7-positive cells to evaluate MSC purity
MSC proliferation
To evaluate the proliferation capacity of MSCs isolated using different digestion enzymes, cells were cultured in growth media for 4 days, and proliferation was quantified using the MTS assay. Microscopic examination revealed no significant differences in cell morphology across enzyme treatments, indicating consistent phenotypic characteristics regardless of the enzyme used (Fig. 3A). Quantitatively, MSCs isolated with TDzyme CT demonstrated the highest proliferation rate, approximately 11% higher than that of the Pronase control group. TDzyme C also enhanced MSC proliferation, showing an 8% increase compared to the control. In contrast, MSCs isolated with TDzyme T exhibited a notable 15% reduction in proliferation, suggesting a negative impact of this enzyme on cell growth potential (Fig. 3B).Fig. 3. Proliferation capacity of MSCs isolated using crude and recombinant proteolytic enzymes. (A) Morphological assessment of MSCs after 4 d of culture, observed under a microscope. (B) Quantitative analysis of proliferation rates using the MTS assay after 4 d of culture
MSCs differentiation
To assess differentiation capacity, MSCs isolated using different enzymatic treatments were cultured in differentiation media for 4 days. Microscopic evaluation of myotube morphology revealed that MSCs isolated using most enzymes underwent successful cell fusion, forming elongated and multinucleated myotubes. However, in the case of TDzyme T, no clear myotube structures were observed, indicating a possible defect in cell fusion ability or a diminished potential for terminal differentiation.
To further clarify the differentiation outcomes, the myotube structure was visualized through immunocytochemistry targeting MYH, a key component of mature myofibrils (green: MYH, blue: nuclei). MSCs isolated with Pronase and TDzyme CT displayed well-formed, elongated, and aligned myotubes, indicating successful differentiation. In contrast, MSCs isolated with TDzyme C and TDzyme T showed disorganized, fragmented structures with poorly defined myotubes, further supporting their reduced differentiation capacity (Fig. 4A).Fig. 4. Differentiation potential of MSCs isolated using crude and recombinant proteolytic enzymes. (A) Bright-field microscopy images showing myotube formation after 4 d in differentiation media. MYH expression (green) was visualized by immunocytochemistry; nuclei were counterstained with DAPI (blue). (B) Creatine kinase activity was measured to assess myogenic differentiation efficiency. (C–E) Gene and protein expression levels of key myogenic markers—MYOD (early), MYOG (intermediate), and MYH (late)—were analyzed using real-time RT-PCR and Western blotting, respectively
Quantitative assessment of differentiation was performed by measuring CK activity, a muscle-specific enzyme that exhibits increased expression during myogenic differentiation. MSCs isolated using TDzyme CT exhibited a 7% higher CK activity compared to those isolated with Pronase, suggesting enhanced differentiation efficiency. In contrast, MSCs isolated with TDzyme C and TDzyme T showed markedly reduced CK activity (9% and 36% lower than Pronase, respectively), aligning with their abnormal myotube morphology and indicating impaired differentiation potential in these groups (Fig. 4B).
To gain deeper insight into the molecular mechanisms underlying these observations, the expression of key myogenic markers was analyzed at both gene and protein levels. The expression of MYOD, an early-stage myogenic regulatory factor, was comparable between MSCs isolated with TDzyme CT and those with Pronase at the gene level; however, protein expression was slightly reduced in the TDzyme CT group. Notably, both TDzyme C and TDzyme T groups showed significantly lower MYOD gene and protein expression than Pronase (Fig. 4C), consistent with their impaired early differentiation signaling.
The expression of MYOG, a mid-stage myogenic differentiation marker, revealed a different trend. MSCs isolated with TDzyme CT exhibited significantly increased MYOG expression at both the gene and protein levels compared to Pronase, suggesting enhanced progression through the myogenic program. Conversely, TDzyme C and TDzyme T groups showed MYOG expression levels similar to those of the Pronase group (Fig. 4D).
Finally, the expression of MYH, a late-stage differentiation marker essential for myofibril assembly, was evaluated. All enzyme groups, except for TDzyme T, demonstrated increased MYH gene and protein expression compared to Pronase. The most pronounced upregulation was observed in the TDzyme CT group, which significantly enhanced both gene and protein levels of MYH, consistent with its superior myotube morphology and CK activity. In contrast, the TDzyme T group not only failed to enhance MYH expression but exhibited lower gene and protein levels than the Pronase group, corroborating its diminished differentiation capacity (Fig. 4E).
Discussion
This study investigated the effects of various proteolytic enzymes (both crude and recombinant) on the isolation, proliferation, and differentiation of chicken MSCs, with a particular focus on their role in promoting myogenic differentiation. The findings provide valuable insights into how the choice of enzyme impacts each stage of the MSC processing workflow, from isolation and purification to proliferation and differentiation.
In terms of tissue digestion efficiency, crude Pronase demonstrated superior muscle digestion capability. This is likely due to its heterogeneous composition, which includes a broad spectrum of proteolytic enzymes such as serine proteases, metalloproteases, aminopeptidases, and carboxypeptidases produced by Streptomyces griseus (Narahashi et al. 1968). These enzymes act synergistically to degrade ECM components, thereby facilitating the release of MSCs. In contrast, recombinant collagenase (TDzyme C) and recombinant thermolysin (TDzyme T), when used individually, exhibited lower tissue digestion efficiency, resulting in reduced MSC yields. This limited efficacy likely stems from the narrow enzymatic profiles of these recombinant enzymes, which restrict their ability to effectively dissociate complex tissue structures.
To overcome the limitations of single recombinant enzymes, TDzyme C and TDzyme T were combined (TDzyme CT). This approach significantly improved MSC yield, cell viability, and myogenic potential, even outperforming Pronase in some aspects. The combination likely benefited from the complementary activities of TDzyme C, which selectively cleaves collagen, and TDzyme T, which non-selectively digests proteins, thus achieving a more balanced and efficient tissue dissociation process.
In terms of MSC purity, the expression of PAX7, a well-established marker for quiescent and activated MSCs (Bosnakovski et al. 2008), remained above 90% for cells isolated using TDzyme CT, similar to levels observed with Pronase. This indicates that the combined recombinant enzymes maintain a high degree of specificity for MSCs, validating their utility for generating pure MSC populations.
Proliferation analysis revealed further distinctions among the enzymes. TDzyme C, when used alone, supported significantly higher MSC proliferation rates than Pronase. This may be due to its collagen-specific activity, which minimizes non-target damage to cell surface proteins critical for proliferation (Eckhard et al. 2013). In contrast, TDzyme T alone led to an approximately 15% decrease in proliferation, likely due to its non-specific proteolytic activity that may compromise membrane integrity or release inhibitory factors during isolation. The highest proliferation rates were observed with TDzyme CT, suggesting that the synergistic enzymatic action created a more favorable environment for subsequent cell growth. These findings underscore the benefit of combining collagen-specific and broader-spectrum proteolytic activities to enhance both isolation efficiency and cell viability.
When evaluating MSC differentiation, TDzyme CT once again outperformed Pronase. MSCs isolated using TDzyme CT exhibited thicker and more defined myotubes under fluorescence microscopy, and they exhibited significantly higher CK activity—a key biochemical marker of myogenic differentiation—compared to those isolated with Pronase. This suggests that not only does TDzyme CT preserve the differentiation capacity of MSCs, it may even enhance it.
In contrast, when TDzyme C and TDzyme T were used individually, CK activity was reduced by 9% and 36%, respectively, compared to Pronase, aligning with the observed morphological defects in myotube formation. These findings reinforce the conclusion that the combination of recombinant enzymes offers a superior alternative to single-enzyme approaches for preserving and enhancing differentiation potential.
This conclusion is further supported by molecular analysis of myogenic regulatory factors. Expression of MYOD (early), MYOG (intermediate), and MYH (late) differentiation markers was comparable to or higher in the TDzyme CT group than in the Pronase group at both gene and protein levels. Most notably, MYH expression, which is essential for myofibril assembly, was significantly upregulated in the TDzyme CT group, consistent with its superior myotube morphology and CK activity. Conversely, TDzyme T resulted in reduced MYH expression, further confirming its limited effectiveness in supporting terminal differentiation.
In terms of the in-depth mechanisms by which these enzymes influence cell growth and differentiation, Pronase is a crude enzyme mixture containing a variety of proteases, including collagenase, with broad-spectrum activity. However, due to its undefined composition, it is difficult to precisely determine its effects on specific membrane proteins, which may compromise cell surface integrity. In contrast, TDzyme C (collagenase-based) and TDzyme T (thermolysin-based) are recombinant enzymes that allow for more selective proteolytic activity, targeting extracellular matrix proteins with higher specificity. When used individually, however, they may exhibit limited efficiency in tissue dissociation. TDzyme CT, a recombinant blend of TDzyme C and TDzyme T, offers a balanced approach—enabling effective degradation of extracellular matrix components while minimizing damage to membrane proteins. This enzymatic synergy contributes to enhanced MSC proliferation and differentiation by preserving cellular integrity and functionality during isolation.
In summary, the combined recombinant proteolytic enzymes TDzyme CT demonstrated improved performance over the crude Pronase in terms of MSC isolation yield, purity, proliferation, and differentiation capacity. These findings position TDzyme CT as a promising, well-defined alternative to traditional crude enzymes for the efficient and effective isolation and expansion of MSCs in avian species.
Ultimately, TDzyme CT demonstrated superior performance in the isolation, proliferation, and differentiation of chicken MSCs, with the added advantage of safety assurance as a recombinant enzyme. These characteristics are expected to enhance the production efficiency and ensure the food safety of chicken cultured meat in future applications.
Conclusion
This study demonstrates that recombinant enzymes, specifically recombinant collagenase and thermolysin, offer significant advantages over traditional crude enzymes in the extraction of chicken MSCs. Notably, the combination of these two recombinant enzymes markedly enhanced MSC isolation, proliferation, and differentiation compared to either enzyme used alone. These findings suggest that recombinant collagenase and thermolysin represent effective and reliable alternatives to crude proteases for obtaining high-purity, high-quality, and functionally active MSCs. This advancement has important implications for muscle biology research and emerging applications such as cultured meat production.
Supplementary Information
Below is the link to the electronic supplementary material.Supplementary file1 (DOCX 17 KB)