Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
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
Cardiac differentiation strategy using a chemically defined medium consisting of just three components: the basal medium RPMI1640, l-ascorbic acid 2-phosphate and rice-derived recombinant human albumin. this protocol produced contractile sheets of up to 95% TNNT2+ cardiomyocytes at a yield of up to 100 cardiomyocytes for every input pluripotent cell. They first assessed chemically defined pluripotent culture on other defined matrices: rh E-cadherin, rh vitronectin, laminin-521, iMatrix-511, human fibronectin and a fibronectin mimetic. Laminin-based matrices resulted in higher growth rates compared to the vitronectin peptide. Fibronectin-based matrices did not support pluripotent growth. All five suitable matrices supported efficient differentiation in CDM3 but only the laminin-based matrices maintained long-term adhesion (>15 d) during CDM3 cardiac differentiation. The authors state that laminin-521 is an optimal matrix for chemically defined differentiation of human iPSC to cardiomyocytes but they still performed all subsequent characterization on the vitronectin peptide due to cost awareness.
Efficient differentiation of human pluripotent stem cells into cardiomyocytes on cell sorting thermoresponsive surface
Sung T.-C., Su H.C, Ling Q.-D., Kumar S.S., Chang Y., Hsu S.T-., Higuchi A.Biomaterials, 2020
The current differentiation process of human pluripotent stem cells (hPSCs) into cardiomyocytes to enhance the purity of hPSC-derived cardiomyocytes requires some purification processes, which are laborious processes. Here, the authors have developed cell sorting plates, which are prepared from coating thermoresponsive poly(N-isopropylacrylamide) and extracellular matrix proteins vitronectin or Biolaminin 521. The Biolaminin 521-coated surface exhibited higher beating colony numbers than the vitronectin-coated surface. This is explained by the fact that hPSC-derived cardiomyocytes express less integrin αVβ5 but more α6β1, where the main binding sites of rVN and LN-521 are integrin αVβ5 and integrin α6β1, respectively. After hPSCs were induced into cardiomyocytes on the thermoresponsive surface coated with Biolaminin 521 for 15 days, the cells were detached partially from the thermoresponsive surface. The detached cells exhibited a higher cardiomyocyte marker of cTnT than the remaining cells on the thermoresponsive surface as well as the cardiomyocytes after purification using conventional cell selection. The detached cells expressed several cardiomyocyte markers, such as α-actinin, MLC2a, and NKX2.5. This study a promising method for the purification of hPSC-derived cardiomyocytes without conventional laborious processes.
Highly sensitive droplet digital PCR method for detection of residual undifferentiated cells in cardiomyocytes derived from human pluripotent stem cells
Kuroda T., Yasuda S., Matsuyama S., Tano K., Kusakawa S., Sawa Y., Kawamata S., Sato Y. Regenerative Therapy, 2015
Human iPSCs were maintained on laminin-521 in Essential 8 medium. Cardiac differentiation on laminin-521 and Matrigel. Adult human cardiomyocytes (Promocell, Heidelberg) were cultured on laminin-211 in the Promocell myocyte growth medium. The authors have established a sensitive assay for the detection of the residual undifferentiated hiPSCs in cardiomyocytes, using droplet digital PCR (ddPCR). LIN28 was the most sensitive marker of residual undifferentiated cells in hiPSC-derived cardiomyocytes but also in other tissues such as liver, heart, pancreas, kidney, spinal cord, corneal epithelium, and lung. Hence, the LIN28/ddPCR method is applicable to the quality control of hiPSC-derived cell therapy products.
Pluripotent stem cell-derived cardiovascular progenitors differentiated on laminin 221 regenerate and improve function of infarcted swine hearts
Yap L, Chong LY, Tan C, Adusumalli S, Seow M, Guo J, Cai Z, Loo SJ, Lim E, Lath N, Ye L, Petretto E, Tryggvason K. BiorXiv, 2021
The authors report a highly reproducible, chemically defined and fully humanized differentiation method of hESCs for the generation of potent cardiovascular progenitors (CVPs), using laminin-221 (Biolaminin 221) as the culture matrix. Transplantation of the CVPs into the myocardial infarcted pig hearts yielded maturation of the progenitor cells to cardiomyocytes and improved cardiac function using only 200 million CVPs, without fatal ventricular arrhythmia occurring. The results may have a significant impact on regenerative cardiology and the method is intended to be used in stem cell therapy of myocardial infarction in humans.
Extracellular Matrix and Integrins in Embryonic Stem Cell Differentiation
Wang H., Luo X. Leighton J.Biochemistry Insights, 2015
Here, the author summarizes the role of the ECM and integrins in the formation of three germ layer progenies. Various ECM–integrin interactions were found, facilitating differentiation toward definitive endoderm, hepatocyte-like cells, pancreatic beta cells, early mesodermal progenitors, cardiomyocytes, neuro-ectoderm lineages, and epidermal cells, such as keratinocytes and melanocytes.
A Chemical Probe that Labels Human Pluripotent Stem Cells
Hirata N., Nakagawa M., Fujibayashi Y., Yamauchi K., Murata A., Minami I., Tomioka M., Kondo T., Kuo T-F., Endo H., Inoue H., Sato S., Ando S., Kawazoe Y., Aiba K., Nagata K., Kawase E., Chang Y-T., Suemori H., Eto K., Nakauchi H., Yamanaka S., Nakatsuji N., Ueda K., Uesugi M. Cell Reports, 2014
The Yamanaka group uses cardiac-specific laminin-211 as the matrix to differentiate iPS cells to cardiomyocytes in a biorelevant environment specific to heart cells. This thus shows that you can use laminin-211 as a cardiac matrix. 326 fluorescent compounds screened to identify a fluorescent probe that is selective for human pluripotent stem cells compared to differentiated cells. hiPSCs were cultured on 3.5 cm culture dishes coated with human laminin 211 and cardiac differentiation was carried out. Cardiac colonies were harvested on day 15 and cultured for 7–10 days in floating culture. A majority of the prepared cells expressed the cardiac markers: cardiac troponin T, a-actinin, and NKX2.5.
iPSC-derived human cardiac progenitor cells improve ventricular remodelling via angiogenesis and interstitial networking of infarcted myocardium
Ja K.P., Miao Q., Zhen Tee N.G., Lim S.Y., Nandihalli M, Ramachandra C.J.A, Mehta A, Shim W. J Cell Mol Med. 2015
Here they investigate the effects of myocardial transplantation of human induced pluripotent stem cell (iPSC)-derived progenitors and cardiomyocytes into acutely infarcted myocardium in immune deficient mice. iPSC cultured on Matrigel and differentiated in EB structures. Differences in integrin and laminin expression between cardiac progenitors and cardiomyocytes were observed. The a6 integrin was higher expressed in progenitors and integrin a1, a2, a3, a7 and b1 higher expression in cardiomyocytes. They observed a distinct swish in expression profile of laminin subunits during cardiac differentiation with laminin-411/421 pre-dominantly expressed early in progenitors and laminin-211/221 expressed later in cardiomyocytes. Improvements of myocardial function in the progenitor group corresponded to increased vascularization and coincided with augmented networking of cardiac telocytes in the interstitial space of the infarcted zone. Laminin-221/211-expressing cardiomyocytes only retained and engrafted around myofibres in the peri-infarct region.