Here is a selection of publications where different laminin isoforms were used to create more authentic cell culture systems.
Laminin as a Potent Substrate for Large-Scale Expansion of Human Induced Pluripotent Stem Cells in a Closed Cell Expansion System
Fernanda C. Paccola Mesquita, Camila Hochman-Mendez, Jacquelynn Morrissey, Luiz C. Sampaio, Doris A. Taylor. Stem Cells Int, 2019
The authors describe the use Quantum Cell Expansion System (QES) as an iPSC monolayer-based expansion system. Human iPSCs were expanded (up to 14-fold) using the QES on two different coatings (laminin 521 (LN521) and vitronectin (VN)). The esults demonstrated that the QES provides the necessary environment for exponential iPSC growth only when LN521 was used. They conclude that the system provides a promising platform to provide the number of cells necessary to recellularize small human-sized organ scaffolds for clinical purposes.
Large-scale production of megakaryocytes in microcarrier supported stirred suspension bioreactors
Eicke D., Baigger A., Schulze K., Latham S.L., Halloin C., Zweigerdt R., Guzman C.A., Blasczyk R., Figueiredo C.Scientific reports, 2018
Although extensive research has been dedicated to developing processes for differentiating PLTs from MKs, no sustainable process for large-scale MK production is available. This study aimed at developing an effective, xeno-free and scalable system to produce high numbers of MKs. In particular, microcarrier beads-assisted stirred bioreactors were evaluated as a means of improving MK yields. This method resulted in the production of 18.7 Å~ 107 MKs per 50 ml medium. Biolaminin 521-coated microcarriers increased MK production per iPSC by up to 10-fold. MKs obtained in this system showed typical features of mature MKs and were able to produce PLTs in vitro and in vivo. To increase safety, MKs produced in the bioreactors were irradiated; a procedure that did not affect their capability to form proPLTs and PTLs after transfusion.
A Scalable Suspension Platform for Generating High-Density Cultures of Universal Red Blood Cells from Human Induced Pluripotent Stem Cells
Jaichandran Sivalingam, Yu SuE, Zhong Ri Lim, Shaul Reuveny, Benoit Malleret, Steve K.W. Oh. Stem Cells Reports, 2020
This article describes a scalable suspension agitation culture platform for the differentiation of human induced pluripotent stem cell-microcarrier aggregates into functional red blood cells. The report describes the cell quantity and quality in culture sizes ranging from 6-well plates to 500 ml spinner flasks, reaching up to 17 million high-quality cells per ml. The process could find applications in future large-scale red blood cell production in controlled bioreactors.
Human Pluripotent Stem Cell Culture: Considerations for Maintenance, Expansion, and Therapeutics
Kevin G. Chen K.G., Mallon B.S., McKay R.D.G., Robey P.GCell stem cell, 2014
In this review, the authors look at different large-scale hPSC culture growth components by comparing cell culture methods (matrices, media, etc.) and identifying the advantage and disadvantages and pitfalls associated with each one. Since laminin-521 was not available when the review was written, they only mention laminin-511 where the cells are seeded as clumps (Rodin et al 2010). The only disadvantage mentioned regarding laminin-511 is that the use of laminin for maintenance and expansion will be to thigh due to the price of the laminins.
Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells
Plaza Reyes A., Petrus-Reurer S. Padrell Sánchez S., Kumar P., Douagi I, Bartuma H., Aronsson M., Westman S., Lardner E., André H Falk A., Nandrot E.F., Kvanta A., Lanner F.Nature Communications, 2020
Here, the authors have performed a comprehensive antibody screening and identify cell surface markers for RPE cells. They identified CD140b, CD56, GD2, and CD184 as central cell surface markers to evaluate hPSC-RPE differentiation efficiency, as well as a potential tool for the enrichment of hPSC-RPE during and after differentiation. They show that these markers can be used to isolate RPE cells during in vitro differentiation and to track, quantify and improve differentiation efficiency. Using these markers together with single-cell RNA-sequencing to evaluate the differentiation process, they have established an efficient, robust, direct and scalable xeno-free and defined monolayer differentiation protocol, where culture on supportive human recombinant Biolaminin 111 and 521 eliminates the need for manual selection, allowing large-scale production of pure hPSC-RPE.