Biology subjects

Jara, C. P.

Publications and source records attributed to Jara, C. P..

4 recordsLinked to original sources

bioRxiv2021

Isolating and Cryo-Preserving Pig Skin Cells for Single Cell RNA Sequencing Study

The Pigskin architecture and physiology are similar to these of humans. Thus, the pig model is valuable for studying skin biology and testing therapeutics for skin diseases. The single-cell RNA sequencing technology allows quantitatively analyzing cell types, cell states, signaling, and receptor-ligand interactome at single-cell resolution and at high throughput. scRNA-Seq has been used to study mouse and human skins. However, studying pigskin with scRNA-Seq is still rare. Here we described a robust method for isolating and cryo-preserving pig single cells for scRNA-Seq. We showed that pigskin could be efficiently dissociated into single cells with high cell viability using the Miltenyi Human Whole Skin Dissociation kit and the Miltenyi gentleMACS Dissociator. Also, we showed that the subsequent single cells could be cryopreserved using DMSO without causing additional cell death, cell aggregation, or changes in gene expression profiles. Using the developed protocol, we were able to identify all the major skin cell types. The protocol and results from this study will be very valuable for the skin research scientific community.

cell biology↗

bioRxiv2020

Glutamic acid promotes hair growth in mice

Glutamic Acid is the main excitatory neurotransmitter in neurons. Abnormal distributions of the glutamic acid receptors have been shown in hyper proliferative models such as psoriasis and skin regeneration. However, the biological function of glutamic acid in the skin remains unclear. Using ex vivo, in vivo and in silico approaches, we showed for the first time that exogenous glutamic acid promotes hair growth and keratinocyte proliferation. Topical application of glutamic acid decreased expression of genes related to apoptosis signaling in the skin. Also, we showed Glutamic acid increased viability and proliferation in cultured human keratinocyte. For the first time, we identified the excitotoxic GA concentration and we provided evidence for the existence of a novel skin signaling pathway mediated by a neurotransmitter controlling keratinocyte and hair follicle proliferation. In perspective, we anticipate our results could be the starting point to elucidate how exogenous glutamic acid from food intake or even endogenous GA from neuropsychiatric disorders modulate skin diseases.

pharmacology and toxicology↗

bioRxiv2020

A smartphone app for individual anesthetic calculation decreased anesthesia-related mortality in mice

Annually, millions of animals are used for experimental purposes. Despite the recommended anesthetic doses being well-known worldwide, the final amounts applied to mice could be different than those calculated. Here, we developed, tested, and validated a mobile app where researchers and operators were able to use personal devices to process body weight, calculate a master anesthetic cocktail, and then apply the individual volume to each mouse. Our objective was to refine anesthesia procedures using information technologies. Our data showed that the "Labinsane" mobile app decreased anesthetic-related deaths upon using weight-adjusted doses of ketamine and xylazine. Also, we validated that the Labinsane mobile app matched all calculations of anesthetic doses. To our knowledge, this is the first report with hundreds of anesthetized mice records and validation and implementation of a mobile app to solve an old but transversal challenge for researchers working with experimental mice.

pharmacology and toxicology↗

bioRxiv2020

Three-Dimensional (3D) Fibronectin Nano-Array Presented on Fibrin Matrix Accelerates Mice Skin Wound Healing

Plasma fibrinogen (F1) and fibronectin (pFN) polymerize to form a fibrin clot that is both a hemostatic and provisional matrix for wound healing. About 90% of plasma F1 has a homodimeric pair of {gamma} chains ({gamma}{gamma}F1) and 10% has a heterodimeric pair of {gamma} and more acidic {gamma} chains ({gamma}{gamma}F1). We have synthesized a novel fibrin matrix exclusively from a 1:1 (molar ratio) complex of {gamma}{gamma}F1 and pFN in the presence of highly active thrombin and recombinant Factor XIII (rFXIIIa). In this matrix, the fibrin nanofibers were wrapped with periodic 200-300 nm wide pFN nanobands (termed {gamma}{gamma}F1:pFN fibrin). In contrast, fibrin made from 1:1 mixture of {gamma}{gamma}F1 and pFN formed a sporadic distribution of "pFN droplets" (termed {gamma}{gamma}F1 +pFN fibrin). The {gamma}{gamma}F1:pFN fibrin enhanced the adhesion of primary human umbilical vein endothelium cells (HUVECs) relative to the {gamma}{gamma}F1+FN fibrin. Three dimensional (3D) culturing showed that the {gamma}{gamma}F1:pFN complex fibrin matrix enhanced the proliferation of both HUVECs and primary human fibroblasts. HUVECs in the 3D {gamma}{gamma}F1:pFN fibrin exhibited a starkly enhanced vascular morphogenesis while an apoptotic growth profile was observed in the {gamma}{gamma}F1 +pFN fibrin. Relative to {gamma}{gamma}F1 +pFN fibrin, mouse dermal wounds that were sealed by {gamma}{gamma}F1:pFN fibrin exhibited accelerated and enhanced healing. This study suggests that a 3D pFN nano-array presented on a fibrin matrix can promote wound healing.

bioengineering↗