In vitro 3D culture is an important model for tissues in vivo. Cells in different locations of 3D tissues are physiologically different, because they are exposed to different concentrations of oxygen, nutrients, and signaling molecules, and to other environmental factors (temperature, mechanical stress, etc). The majority of high-throughput assays based on 3D cultures, however, can only detect the average behavior of cells in the whole 3D construct. Isolation of cells from specific regions of 3D cultures is possible, but relies on low-throughput techniques such as tissue sectioning and micromanipulation. Based on a procedure reported previously (“cells-in-gels-in-paper” or CiGiP), this paper describes a simple method for culture of arrays of thin planar sections of tissues, either alone or stacked to create more complex 3D tissue structures. This procedure starts with sheets of paper patterned with hydrophobic regions that form 96 hydrophilic zones. Serial spotting of cells suspended in extracellular matrix (ECM) gel onto the patterned paper creates an array of 200 micron-thick slabs of ECM gel (supported mechanically by cellulose fibers) containing cells. Stacking the sheets with zones aligned on top of one another assembles 96 3D multilayer constructs. De-stacking the layers of the 3D culture, by peeling apart the sheets of paper, “sections” all 96 cultures at once. It is, thus, simple to isolate 200-micron-thick cell-containing slabs from each 3D culture in the 96-zone array. Because the 3D cultures are assembled from multiple layers, the number of cells plated initially in each layer determines the spatial distribution of cells in the stacked 3D cultures. This capability made it possible to compare the growth of 3D tumor models of different spatial composition, and to examine the migration of cells in these structures.
This paper describes the development of MEMS force sensors constructed using paper as the structural material. The working principle on which these paper-based sensors are based is the piezoresistive effect generated by conductive materials patterned on a paper substrate. The device is inexpensive (~$0.04/device for materials), simple to fabricate, light weight, and disposable. Paper can be readily folded into three-dimensional structures to increase the stiffness of the sensor while keeping it light in weight. The entire fabrication process can be completed within one hour without expensive cleanroom facilities using simple tools (e.g., a paper cutter and a painting knife). We demonstrated that the paper-based sensor can measure forces with moderate performance (i.e., resolution: 120 µN, measurement range: ±16 mN, and sensitivity: 0.84 mV/mN), and we applied this sensor to characterizing the mechanical properties of a soft material. Leveraging the same sensing concept, we also developed a paper-based balance with a measurement range of 15 g, and a resolution of 390 mg.2
Increasing evidence supports the importance of the breast milk microbiome in seeding the infant gut. However, the origin of bacteria in milk and the process of milk microbe-mediated seeding of infant intestine need further elucidation. Presumed sources of bacteria in milk include locations of mother-infant and mother-environment interactions. We investigate the role of mother-infant interaction on breast milk microbes. Shotgun metagenomics and 16S rRNA gene sequencing identified milk microbes of mother-infant pairs in breastfed infants and in infants that have never latched. Although breast milk has low overall biomass, milk microbes play an important role in seeding the infant gut. Breast milk bacteria were largely comprised of Staphylococcus, Streptococcus, Acinetobacter, and Enterobacter primarily derived from maternal areolar skin and infant oral sites in breastfeeding pairs. This suggests that the process of breastfeeding is a potentially important mechanism for propagation of breast milk microbes through retrograde flux via infant oral and areolar skin contact. In one infant delivered via Caesarian section, a distinct strain of Bifidobacteria breve was identified in maternal rectum, breast milk and the infant's stool potentially suggesting direct transmission. This may support the existence of microbial translocation of this anaerobic bacteria via the enteromammary pathway in humans, where maternal bacteria translocate across the maternal gut and are transferred to the mammary glands. Modulating sources of human milk microbiome seeding potentially imply opportunities to ultimately influence the development of the infant microbiome and health.
2021 Increasing evidence supports the importance of the breast milk microbiome in seeding the infant 22 gut. However, the origin of bacteria in milk and the process of milk microbe-mediated seeding of 23 infant intestine need further elucidation. Presumed sources of bacteria in milk include locations 24 of mother-infant and mother-environment interactions. We investigate the role of mother-infant 25 interaction on breast milk microbes. Shotgun metagenomics and 16S rRNA gene sequencing 26 identified milk microbes of mother-infant pairs in breastfed infants and in infants that have never 27 latched. Although breast milk has low overall biomass, milk microbes play an important role in 28 seeding the infant gut. Breast milk bacteria were largely comprised of Staphylococcus, 29 Streptococcus, Acinetobacter, and Enterobacter primarily derived from maternal areolar skin and 30 infant oral sites in breastfeeding pairs. This suggests that the process of breastfeeding is a 31 potentially important mechanism for propagation of breast milk microbes through retrograde flux 32 via infant oral and areolar skin contact. In one infant delivered via Caesarian section, a distinct 33 strain of Bifidobacteria breve was identified in maternal rectum, breast milk and the infant's 34 stool potentially suggesting direct transmission. This may support the existence of microbial 35 translocation of this anaerobic bacteria via the enteromammary pathway in humans, where 36 maternal bacteria translocate across the maternal gut and are transferred to the mammary glands.
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