ABSTRACT
Adipose tissue-derived mesenchymal stromal/stem cells (ASCs) are considered important tools in regenerative medicine and are being tested in several clinical studies. Porcine models are frequently used to obtain adipose tissue, due to the abundance of material and because they have immunological and physiological similarities with humans. However, it is essential to understand the effects and safe application of ASCs from pigs (pASCs) as an alternative therapy for diseases. Although minipigs are easy-to-handle animals that require less food and space, acquiring and maintaining them in a bioterium can be costly. Thus, we present a protocol for the isolation and proliferation of ASCs isolated from adipose tissue of farm pigs. Adipose tissue samples were extracted from the abdominal region of the animals. Because the pigs were not raised in a controlled environment, such as a bioterium, it was necessary to carry out rigorous procedures for disinfection. After this procedure, cells were isolated by mechanical dissociation and enzymatic digestion. A proliferation curve was performed and used to calculate the doubling time of the population. The characterization of pASCs was performed by immunophenotyping and cell differentiation in osteogenic and adipogenic lineages. The described method was efficient for the isolation and cultivation of pASCs, maintaining cellular attributes, such as surface antigens and multipotential differentiation during in vitro proliferation. This protocol presents the isolation and cultivation of ASCs from farm pig as an alternative for the isolation and cultivation of ASCs from minipigs, which require strictly controlled maintenance conditions and a more expensive process.
ABSTRACT
Standard formalin embalming is the most important of all work with cadavers in anatomy laboratories, as it keeps the tissue strict, insoluble and protected against deterioration. The most commonly used substance for preservation of cadavers and tissues is formaldehyde, a preservative because it is inexpensive, easy penetration and fast action on the parts. Another substance used is glycerin with rapid action and dehydrating fixative, used also for the preservation of anatomical specimens. This study aimed to compare two techniques that use glycerin in conserving parts of the central nervous system of animals. We evaluated the properties and fixing of conservative solutions applied. The two techniques chosen for this work were: the Giacomini and Torres method. In the Torres method, the tissue was more flexible and easy to visualize the structures. In the Giacomini method, the tissues were dark colored, rigid, with little flexibility. This technique requires a shorter time of immersion, compared to Torres. We conclude that the most appropriate method for application in laboratories of anatomy and applicability in practical lessons is technique was Torres.