Biorelevant culture of cardiac cells on Biolaminin 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
Highy reproducible, chemically defined, xeno-free laminin-based differentiation protocol to generate stem cell-derived cardiovascular progenitors (CVPs) was reported in 2019 (Yap, 2019). 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.
With the use of cardiac muscle-specific laminin-211, human iPSCs were effectively 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., demonstrating 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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Laminin 521 maintains differentiation potential of mouse and human satellite cell-derived myoblasts during long-term culture expansion
Penton C.M., Badarinarayana V., Prisco J., Powers E., Pincus M., Allen R.E., August P.R. Skeletal Muscle, 2016
Instructions: Coating with Biolaminin substrates
Protocol and concentration calculations for coating cultureware with Biolaminin
Chemically defined generation of human cardiomyocytes
Burridge P., Elena Matsa E., Shukla P., Lin Z., Churko J., Ebert A., Lan F., Diecke S., Huber B., Mordwinkin N., Plews J., Abilez O., Cui B., Gold J. & Wu J. Nature methods, 2014
Biolaminin Key Advantages
Biolaminin 521 is an optimal matrix for chemically defined differentiation of human iPSC to cardiomyocytes. Laminin-521 supports increased proliferation during expansion and superior differentiation with myotube hypertrophy, larger myotubes and higher amounts of nuclei per myotube. Biolaminin 211 can effectively be used for cardiac differentiation of human iPSC and also for the culture of adult human cardiomyocytes.
Specific laminin isoforms are present in different tissue microenvironments and are essential for cell survival, proliferation, and differentiation. Biolaminin products allow you to imitate the natural cell-matrix interactions in vitro.
Our products have consistent composition and quality. This enables minimized variability between experiments and uniform pluripotency gene expression profiles between different cell lines.
All our matrices are chemically defined and animal origin-free, which makes them ideal substrates for each level of the scientific process – from basic research to clinical applications.
Numerous scientists have found our products and finally succeeded in their specific stem cell application. The power of full-length laminins incorporated into various cell systems is well documented in scientific articles and clinical trials.
Biolaminin 521 LN (LN521)
Human recombinant laminin 521Biolaminin 521 LN is the natural laminin for pluripotent stem cells and therefore reliably facilitates self-renewal of human ES and iPS cells in a chemically defined, feeder-free and animal origin-free stem cell culture system. LN521 is animal origin-free to the primary level.VIEW product
Biolaminin 221 LN (LN221)
Human recombinant laminin 221Biolaminin 221 supports the growth, survival, and differentiation of a wide range of tissue-specific cell types, including cardiac cells and skeletal muscle cells.VIEW product
Biolaminin 211 LN (LN211)
Human recombinant laminin 211Biolaminin 211 supports the growth, survival, and differentiation of a wide range of tissue-specific cell types, including motor neurons, cardiac cells, and skeletal muscle cells.VIEW product
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