PRODUCT RECOMMENDATION
LAMININ-521, LAMININ-211 and LAMININ-221
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PROTOCOL
Long-term expansion and differentiation of myogenic progenitors on laminin
CUSTOMER TESTIMONIALS
"Laminin-521 shifts away from the paradigm that muscle cells lose their ability to differentiate after culturing. Muscle differentiation on laminin-521 is nothing short of spectacular, yielding increased numbers of nuclei per myotube and improved myotube organization. Surprisingly, muscle cells expanded on laminin-521 maintain their ability to differentiate into myotubes after long-term passaging, better-resembling differentiation of freshly isolated cells."
Dr. Christopher Penton, Icagen, Inc., USA
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ARTICLES
HOW TO CULTURE CARDIAC CELLS ON LAMININ SUBSTRATES
Laminin expression in muscle tissue
Laminin-211 and laminin-221 are expressed specifically in the basal lamina of striated muscles and an expression deficiency in the alpha-2 laminins results in muscular dystrophy, accompanied by a dilated cardiomyopathy (Oliviéro, 2000). Laminin isoforms containing the alpha-2, -4 and -5 subunits are critically important for the maintenance and development of heart muscle tissue (Miner, 1997). During stem cell, cardiac differentiation, a distinct swish in the expression profile of laminin subunits have been reported with laminin-411/421 pre-dominantly expressed early in progenitors and laminin-211/221 expressed later in cardiomyocytes (Ja, 2015). In the adult heart, the main laminin isoforms expressed are laminin-211 and laminin-221 but laminin-521 has also shown to be expressed.
Effective cardiomyocyte differentiation on cardiac-specific laminin isoforms
In a Cell Reports article from 2019 (Yap, 2019), the authors present a high reproducible, chemically defined, xeno-free laminin-based differentiation protocol to generate stem cell-derived cardiovascular progenitors (CVPs). Laminin-221 (LN-221) was identified as the most likely expressed cardiac laminin and the authors show that this protein promotes differentiation of pluripotent human embryonic stem cells (hESCs) toward cardiomyocyte lineage and downregulates pluripotency and teratoma-associated genes. Single-cell RNA sequencing of CVPs derived from hESC lines identified three main progenitor subpopulations. These CVPs were transplanted into myocardial infarction mice, where heart function was improved as measured by echocardiogram and human heart muscle bundle formation was identified histologically.
In a publication by a group of Japanese scientists, they have shown that with the use of cardiac muscle-specific laminin-211, human iPSCs can be effectively be differentiated into cardiomyocytes using small molecules (Minami, 2012). The same researches later published another article with Shinya Yamanaka as co-author, where they again used laminin-211 for cardiac differentiation of human iPSC (Hirata, 2014). Laminin-211 has also been used for the culture of adult human cardiomyocytes (Kuroda, 2015).
Laminin-521 also has properties that improve the isolation and expansion of adult stem cells and progenitors. In a publication by Kuroda et al., iPSCs were efficiently expanded and differentiated to cardiomyocytes on laminin-521 (Kuroda, 2015). Dr. Joseph Wu´s lab also showed that laminin-521 is an optimal matrix for chemically defined differentiation of human iPSC to cardiomyocytes (Burridge, 2014). Laminin-521 was compared to 5 other feeder-free matrices and the authors concluded that:
- Laminin-521 had the fastest human iPSC growth rate
- Laminin-521 generated the highest number of cardiomyocytes
- Only laminin-521 could sustain long-term adhesion (>15 d) during cardiac differentiation in chemically defined medium
This in accordance with recent data presented by researchers at Icagen Inc. demonstrates that the laminin-521 cell culture matrix maintains the differentiation potential of mouse and human satellite cell-derived myoblasts, even during long-term culture expansion (Penton, 2016). Laminin-521 supports increased proliferation during expansion and superior differentiation with myotube hypertrophy, larger myotubes and higher amounts of nuclei per myotube. Moreover, Penton et al. show that laminin-521 supports more consistent and reliable differentiation over long-term culture and is the only substrate facilitating high-level fusion following long-term culture. Laminin-521 supports increased differentiation potential without altering the traditional Pax7/MyoD paradigm and the results are translational across several mouse backgrounds, human cells, and disease states.
Laminin IHC staining of human heart muscle
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Expansion of human PSC
Mesenchymal stem cells
Clonal cell culture applications
Eye cells
Cardiac cells
Neural cells
Skeletal muscle cells
Kidney cells
Hepatic cells
Cancer cells
Lung cells
Animal stem cells
Endothelial cells
Pancreatic cells
Intestinal cells
Normal and cancerous mammary cells
Epithelial cells
Biolaminin 521 CTG
Biolaminin 521 CTG cell therapy grade cell culture matrix makes pluripotent stem cell culture easy. A defined, animal origin-free and biologically relevant cell culture system for better cell models.
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Expansion of human PSC
Clonal cell culture applications
Eye cells
Cardiac cells
Neural cells
Skeletal muscle cells
Kidney cells
Hepatic cells
Cancer cells
Lung cells
Animal stem cells
Mesenchymal stem cells
Endothelial cells
Pancreatic cells
Intestinal cells
Normal and cancerous mammary cells
Epithelial cells
Biolaminin 521 MX
Biolaminin 521 MX research-grade cell culture matrix makes pluripotent stem cell culture easy. A defined, animal origin-free and biologically relevant cell culture system for better cell models.
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Expansion of human PSC
Mesenchymal stem cells
Clonal cell culture applications
Eye cells
Cardiac cells
Neural cells
Skeletal muscle cells
Kidney cells
Hepatic cells
Cancer cells
Lung cells
Animal stem cells
Endothelial cells
Pancreatic cells
Intestinal cells
Normal and cancerous mammary cells
Epithelial cells
Biolaminin 521 LN
Biolaminin 521 LN research-grade cell culture matrix makes pluripotent stem cell culture easy. A defined, animal origin-free and biologically relevant cell culture system for better cell models.