A modified animal-free serum technique for efficient isolation and proliferation of mesenchymal stem cells from Hoffa fat pad.

We focus on this study in designing an alternative technique for obtaining mesenchymal stem cells (MSCs) from residual tissue, Hoffa fat, in arthroscopic procedures. Two males and two females were included, and underwent knee arthroscopy; a sample of infrapatellar adipose tissue was obtained with basket forceps. The primary culture was made using the explant method and the culture media: DMEM-high glucose, supplemented with 10% of inactivated human allogeneic serum. All the cellular cultures remained under culture conditions for three weeks, after that by flow cytometry the cells were characterized by MSCs antibody panel: CD105, CD73 and CD90. Subsequently, in the first pass, the MSCs were cultured in commercial human chondrogenic, osteogenic and adipogenic mediums, respectively. After primary culture, we obtained on average 95,600.00 ± 7,233.26 cells/cm2, and the duplication time of MSCs isolate from Hoffa fat pad was established in 39 hours. By flow cytometry, we found that surface markers percentage for expanded MSCs (CD105, CD73, CD90) in primary culture significantly increased and its morphology was fibroblastic-like. After differentiation culture which was made in the first pass, by immunofluorescence, we obtained positive cell markers for three lineages of differentiation, adipocytes: LPL protein, osteocytes: RUNX2, Osteopontin, chondrocytes: SOX9, Aggrecan and COL2A1. We managed to isolate a significant number of MSCs from this source using an easy method to implement and minimal nutrient supplementation, with high potential for differentiation to mature mesenchymal tissues and potential use in basic experimental, preclinical and even clinical research.

Synovial membrane mesenchymal stem cells for cartilaginous tissues repair.

BACKGROUND: The present review is focused on general aspects of the synovial membrane as well as specialized aspects of its cellular constituents, particularly the composition and location of synovial membrane mesenchymal stem cells (S-MSCs). S-MSC multipotency properties are currently at the center of translational medicine for the repair of multiple joint tissues, such as articular cartilage and meniscus lesions. METHODS AND RESULTS: We reviewed the results of in vitro and in vivo research on the current clinical applications of S-MSCs, surface markers, cell culture techniques, regenerative properties, and immunomodulatory mechanisms of S-MSCs as well as the practical limitations of the last twenty-five years (1996 to 2021). CONCLUSIONS: Despite the poor interest in the development of new clinical trials for the application of S-MSCs in joint tissue repair, we found evidence to support the clinical use of S-MSCs for cartilage repair. S-MSCs can be considered a valuable therapy for the treatment of repairing joint lesions.

Uso y aplicación de la tecnología de impresión y bioimpresión 3D en medicina / Use and application of 3D printing and bioimpression technology in medicine

RESUMEN En el mundo actual, las llamadas "tecnologías de fabricación por adición" o impresión 3D también llamado prototipado rápido, han trascendido las fronteras de casi todos los campos de la ciencia, y su incursión en la medicina es cada vez mayor. Es justamente en el campo médico que esta tecnología de impresión por adición ha evolucionado a la bioimpresión, que incluye un proceso de cultivo celular en laboratorio haciendo posible la formación de órganos y/o tejidos personalizados. Para la impresión tridimensional de órganos en humanos se toman muestras de un tejido o células madre del paciente, para ser cultivadas y expandidas en laboratorio para su posterior diferenciación a una línea celular específica. Para este proceso se utiliza un material sólido como andamio a temperatura ambiente con un punto de fusión conocido. En la creación de un modelo para la fabricación de un órgano o tejido en impresión 3D, se utilizan los estudios de imágenes médicas de los pacientes intentando preservar al máximo la anatomía de las estructuras que se desean reproducir. En este artículo se abordan las bases y el potencial uso de esta tecnología en el área médica.

ABSTRACT In today's world, so-called "addition manufacturing technologies" or 3D printing also called rapid prototyping have transcended the borders of almost every field of science and medicine is no exception. It is not surprising that its exploration for practical uses is increasing. In medicine, this technology of printing by addition has evolved to bioprinting, which occurs by a special process, from cells grown in a laboratory, which makes possible its transformation into a type of organs tailored to the patient. The three-dimensional impression of human organs requires take samples of tissues or stem cells from the patient, which are grown in the laboratory waiting to multiply or differentiate to other cell lines; then, to create said object, a solid material at room temperature and with a known melting point is applied layer by layer. Currently the use of this technology uses the medical images of patients trying to preserve the anatomy of the structures that they want to reproduce. In this article the bases and the potential use of this technology in the medical area will be addressed.

Behavior of multipotent stem cells isolated in mobilized peripheral blood from sheep after culture with human chondrogenic medium.

Today, regenerative medicine requires new sources of multipotent stem cells for their differentiation to chondrocytes using the mediums of differentiation available in the market. This study aimed to determine whether the Mesenchymal Stem Cells (MSCs) isolated from Mobilized Peripheral Blood (MPB) in sheep using the Granulocyte Colony-Stimulating Factor (G-CSF), have the ability of first acquire a fibroblast-like morphology after being forced out of the bone marrow niche by G-CSF and second, if the cells have the capacity to express collagen type-II α I in primary culture using a human commercial media of differentiation. Six Suffolk male sheep with age of 2 years were mobilized using G-CSF. One subcutaneous injection of 10 mcg per kilogram of bodyweight were administered every 24 h during three consecutive days. At day four, a sample of 20 mL of peripheral blood was harvested, afterwards, monocytes cells were separated by ficoll gradient. The mobilized MSCs were expanded in primary culture in DMEM medium supplemented with 10% adult sheep serum for three weeks and characterized by an antibody panel for surface markers: CD105, CD90, CD73, CD34, and CD45, before and after primary culture. Subsequently, an aliquot of cells in the first pass were cultured in a commercial human chondrogenic medium for three weeks. As a result, the percentage of surface markers for MSCs (CD105, CD90, CD73) in expanded cells in primary culture significantly increased, at the same time a decrease in the markers for hematopoietic cells (CD34 and CD45) was observed and the cells morphology was fibroblast-like. After three weeks of differentiation culture, the immunofluorescence analysis evidenced the expression of collagen-type-II. It was concluded that Mesenchymal Stem Cells isolated from mobilized peripheral blood in sheep have the ability to pre-differentiate into chondral like cells and express collagen type-II when are stimulated with a human commercial chondrogenic medium in monolayer culture.