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Sand-mediated ice seeding enables serum-free low-cryoprotectant cryopreservation of human induced pluripotent stem cells

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Bioactive Materials 2021年第12期6卷 4377-4388页

作者： Bin Jiang Weijie Li samantha stewart Wenquan Ou Baolin Liu Pierre Comizzoli Xiaoming He Fischell Department of Bioengineering University of MarylandCollege ParkMD20742USA Institute of Biothermal Technology University of Shanghai for Science and TechnologyShanghai200093China Smithsonian Conservation Biology Institute National Zoological ParkWashingtonDC20008USA Robert E.Fischell Institute for Biomedical Devices University of MarylandCollege ParkMD20742USA Marlene and stewart Greenebaum Comprehensive Cancer Center University of MarylandBaltimoreMD21201USA

Human induced pluripotent stem cells(hiPSCs)possess tremendous potential for tissue regeneration and banking hiPSCs by cryopreservation for their ready availability is crucial to their widespread ***,contemporary methods for hiPSC cryopreservation are associated with both limited cell survival and high concentration of toxic cryoprotectants and/or *** latter may cause spontaneous differentiation and/or introduce xenogeneic factors,which may compromise the quality of ***,sand from nature is discovered to be capable of seeding ice above􀀀10◦C,which enables cryopreservation of hiPSCs with no serum,much-reduced cryoprotectant,and high cell ***,the cryopreserved hiPSCs retain high pluripotency and functions judged by their pluripotency marker expression,cell cycle analysis,and capability of differentiation into the three germ *** unique sand-mediated cryopreservation method may greatly facilitate the convenient and ready availability of high-quality hiPSCs and probably many other types of cells/tissues for the emerging cell-based translational medicine.

关键词： Sand Ice seeding Cryopreservation iPSC Stem cell

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Rock inhibitor may compromise human induced pluripotent stem cells for cardiac differentiation in 3D

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Bioactive Materials 2022年第3期7卷 508-522页

作者： Bin Jiang Wenquan Ou James G.Shamul Hao Chen Sarah Van Belleghem samantha stewart Zhenguo Liu John P.Fisher Xiaoming He Fischell Department of Bioengineering University of MarylandCollege ParkMD20742United States Division of Cardiovascular Medicine University of Missouri School of MedicineColumbiaMO65212United States Marlene and stewart Greenebaum Comprehensive Cancer Center University of MarylandBaltimoreMD21201United States

Cardiomyocytes differentiated from human induced pluripotent stem cells(iPSCs)are valuable for the understanding/treatment of the deadly heart diseases and their drug ***,the very much needed homogeneous 3D cardiac differentiation of human iPSCs is still ***,it is discovered surprisingly that Rock inhibitor(RI),used ubiquitously to improve the survival/yield of human iPSCs,induces early gastrulation-like change to human iPSCs in 3D culture and may cause their heterogeneous differentiation into all the three germ layers(i.e.,ectoderm,mesoderm,and endoderm)at the commonly used concentration(10μM).This greatly compromises the capacity of human iPSCs for homogeneous 3D cardiac *** reducing the RI to 1μM for 3D culture,the human iPSCs retain high pluripotency/quality in inner cell mass-like solid 3D ***,the beating efficiency of 3D cardiac differentiation can be improved to more than 95%in~7 days(compared to less than~50%in 14 days for the 10μM RI condition).Furthermore,the outset beating time(OBT)of all resultant cardiac spheroids(CSs)is synchronized within only 1 day and they form a synchronously beating 3D construct after 5-day culture in gelatin methacrylol(GelMA)hydrogel,showing high homogeneity(in terms of the OBT)in functional maturity of the ***,the resultant cardiomyocytes are of high quality with key functional ultrastructures and highly responsive to cardiac *** discoveries may greatly facilitate the utilization of human iPSCs for understanding and treating heart diseases.

关键词： Episomal iPSC Cardiomyocyte Spheroid GelMA

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Noncovalent reversible binding-enabled facile fabrication of leak-free PDMS microfluidic devices without plasma treatment for convenient cell loading and retrieval

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Bioactive Materials 2022年第10期7卷 346-358页

作者： Bin Jiang Alisa White Wenquan Ou Sarah Van Belleghem samantha stewart James G.Shamul Shaik O.Rahaman John P.Fisher Xiaoming He Fischell Department of Bioengineering University of MarylandCollege ParkMD20742USA Department of Nutrition and Food Science University of MarylandCollege ParkMD20742USA Marlene and stewart Greenebaum Comprehensive Cancer Center University of MarylandBaltimoreMD21201USA

The conventional approach for fabricating polydimethylsiloxane(PDMS)microfluidic devices is a lengthy and inconvenient procedure and may require a clean-room microfabrication facility often not readily ***,living cells can’t survive the oxygen-plasma and high-temperature-baking treatments required for covalent bonding to assemble multiple PDMS parts into a leak-free device,and it is difficult to disassemble the devices because of the irreversible covalent *** a result,seeding/loading cells into and retrieving cells from the devices are ***,we discovered that decreasing the curing agent for crosslinking the PDMS prepolymer increases the noncovalent binding energy of the resultant PDMS surfaces without plasma or any other *** enables convenient fabrication of leak-free microfluidic devices by noncovalent binding for various biomedical applications that require high pressure/flow rates and/or long-term cell culture,by simply hand-pressing the PDMS parts without plasma or any other treatment to bind/*** this method,multiple types of cells can be conveniently loaded into specific areas of the PDMS parts before assembly and due to the reversible nature of the noncovalent bonding,the assembled device can be easily disassembled by hand peeling for retrieving *** with 3D printers that are widely available for making masters to eliminate the need of photolithography,this facile yet rigorous fabrication approach is much faster and more convenient for making PDMS microfluidic devices than the conventional oxygen plasma-baking-based irreversible covalent bonding method.

关键词： Binding energy 3D printing Soft lithography Microfluidics Polydimethylsiloxane

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