Potential Agents against Plasma Leakage

Shock due to severe plasma leakage may happen in infectious diseases such as severe dengue and sepsis due to various bacterial infections, which may be deleterious and may lead to death. Various substances and proteins are known to modulate the effects of proleakage mediators and counteract the deleterious effect of plasma leakage. Some of the various substances and proteins such as focal adhesion kinase (FAK), the Rho GTPases, protein kinase A, and caveolin-1 have dual actions; therefore they are not suitable for therapy. However, sphingosine 1phosphate and its receptor agonists, Angiopoetin-1, Slit, and Bbeta15–42 may be promising

Prospect of Induced Pluripotent Stem Cell Genetic Repair to Cure Genetic Diseases

In genetic diseases, where the cells are already damaged, the damaged cells can be replaced by new normal cells, which can be differentiated from iPSC. To avoid immune rejection, iPSC from the patient's own cell can be developed. However, iPSC from the patients's cell harbors the same genetic aberration. Therefore, before differentiating the iPSCs into required cells, genetic repair should be done. This review discusses the various technologies to repair the genetic aberration in patient-derived iPSC, or to prevent the genetic aberration to cause further damage in the iPSC-derived cells, such as Zn finger and TALE nuclease genetic editing, RNA interference technology, exon skipping, and gene transfer method. In addition, the challenges in using the iPSC and the strategies to manage the hurdles are addressed

Prospect of stem cell conditioned medium in regenerative medicine

Background. Stem cell-derived conditioned medium has a promising prospect to be produced as pharmaceuticals for regenerative medicine. Objective. To investigate various methods to obtain stem cell-derived conditioned medium (CM) to get an insight into their prospect of application in various diseases. Methods. Systematic review using keywords "stem cell" and "conditioned medium" or "secretome" and "therapy. " Data concerning treated conditions/diseases, type of cell that was cultured, medium and supplements to culture the cells, culture condition, CM processing, growth factors and other secretions that were analyzed, method of application, and outcome were noted, grouped, tabulated, and analyzed. Results. Most of CM using studies showed good results. However, the various CM, even when they were derived from the same kind of cells, were produced by different condition, that is, from different passage, culture medium, and culture condition. The growth factor yields of the various types of cells were available in some studies, and the cell number that was needed to produce CM for one application could be computed. Conclusion. Various stem cell-derived conditioned media were tested on various diseases and mostly showed good results. However, standardized methods of production and validations of their use need to be conducted

Next Generation Sequencing as Rapid Diagnosis of Multidrug Resistance Tuberculosis

Multi-drug-resistant tuberculosis (MDR-TB) is a threat to global health. In 2018, TB related death was estimated to be more than 1.5 million cases worldwide. Conventional diagnostic method, which requires a long time to get a result, causes delays in new cases discoveries that lead to delayed therapy. Further, delayed and inadequate therapy causes an increase in the level of resistance to anti-TB drugs that may lead to death. Therefore, diagnostic tools, which can detect quickly and accurately, are highly needed. Early and timely detection is crucial for globally effective TB control, but this is not popular in developing countries, especially in Asia. Therefore, the objective of this review is to provide current information on the use of NGS as a rapid diagnostic tool for MDR-TB, especially in Asian populations, and to highlight the various MDR genes.Keywords: Next Generation sequencing; Multi-Drug Resistance; Tuberculosis; Mycobacterium tuberculosis; Rapid diagnosi

Effect of Platelet Rich Plasma on Post Cryopreservation Viability, Morphology and Proliferation of Human Umbilical Cord Stem Cells

Abstract: In most cryopreservation medium, Fetal Bovine Serum (FBS) is used as supplement, while it is well known that FBS contains xenoproteins that can be incorporated into the cells and may be harmfull, as they can elicit immune response. Therefore, finding other xenofree materials as FBS alternative in cryopreservation medium is very important. Platelet Rich Plasma (PRP) is albumin rich and is a candidate for FBS alternative as cryopreservation supplement. Albumin is a natural extracellular cryoprotective agent that stabilizes impaired cell membrane during cryopreservation. This was an in vitro analytical study to to compare the effect of PRP and FBS as supplement in cryopreservation medium on human umbilical cord stem cells. In this study the stem cells were isolated from an umbilical cord tissue by explant method and propagated untill we got enough cells for cryopreservation. Cryopreservations were done using eight types of protocol, which differed in type and concentration of supplement and cell concentration. The effect of the eight protocols were compared in terms of post cryopreservation cell viability, morphology, cell size and proliferation. There were no difference between FBS and PRP supplemented cryopreservation media in term of cell viability and morphology. PRP supplemented medium showed better post cryopreservation performance in cell size and proliferation. PRP can be used as an alternative to FBS in cryopreservation medium for human umbilical cord tissue derived stem cells

Passage Effect on Aging of Human Umbilical Cord Derived Mesenchymal Stem Cell

To be used in regenerative medicine, cells should be checked for various conditions, including cell aging. This study aimed to learn senescent profile and its relation to cell viability, proliferation and cell size in various passages, which were done in α-MEM-10% PRP medium, until senescence-associated β-galactosidase (SA-β-Gal) positive cells were found. Stem cells were isolated from umbilical cord tissue by multiple harvest explant method, cultured in α-MEM-10% PRP until P-17 and stained using SA-β-Gal staining. Viability, Population Doubling Time (PDT), percentage SA-β-Gal (+) and cell size at 30% confluent and at senescent staining were analyzed. Passages with SA-β-Gal (+) and (-) were compared in term of viability, PDT and cell size at 30% confluent. Further, cell size at senescent staining between SA-β-Gal (+) and (-) groups were compared. Viability and PDT showed no significant difference between SA-β-Gal (+) and (-) groups, while cell size at 30% confluence showed significant increase in SA-β-Gal (+) compared to (-) groups. Further, cell size in senescent staining showed significantly smaller cell size in SA-β-Gal (-) compared to SA-β-Gal (+) cells. Moreover, this study showed that even in SA-β-Gal (+) group, viability was greater than 91%, PDT was less than 2.1 days and cell size was less than 2602 µm2. In conclusion, umbilical cord derived MSCs that were cultured in α-MEM-10% PRP began to undergo aging at P-10. Morphological criteria of UC-MSC aging were cell size greater than 2602 µm2 with a change in morphology toward a rounded shape

Comparison of in vitro Vasculogenesis Potential between ASCs with BM-MSCs

Tube formation assay is the most widely used method as a vasculogenesis/ angiogenesis test in vitro. Mesenchymal stem cells (MSCs) are multipotent adult cells. The paracrine effect of MSCs on neovascularization is well known. In general, MSCs do not express CD34 hematopoietic surface marker, but according to some experts, bone marrow mesenchymal stem cells (BM-MSCs) express CD34 in vivo and lose their expression when they are cultured in vitro, while adipose-derived stem cells (ASCs) still have CD34 expression in the early passages when cultured in vitro. BM-MSCs are the most widely used MSC, but ASCs are also used in stem cell therapy and tissue engineering for angiogenesis purposes. Until now, the potential of vasculogenesis between ASCs and BM-MSCs is still unclear. Expression of CD34 is also unknown whether affecting the quality of tube formation. This study wanted to compare the potential of vasculogenesis between ASC and BM-MSCs through tube formation test and CD34 expression.Measurements of vasculogenesis quality showed higher tube length, number of loopsand mean number of branch points on BM-MSC. Both BM-MSCs and ASCs showed low CD34 levels.BM-MSCs showed better tube formation ability compared with ASCs. No association was found between CD34 levels and MSC vasculogenesis capability

Peningkatan Kadar Imunosupresan IDO (Indoleamin 2,3-dioksigenase) pada Supernatan Kultur Sel Punca Mesenkim yang Distimulasi dengan Agregat Imunoglobulin G

Mesenchymal stem cells have been known to have the nature of supressing immune response. This cell have been proven to reduce the symptoms of organ transplants rejection. Clinically proofing is also being performed on an autoimmune disease caused by the complex immune such as systemic lupus erythematosus (SLE). This study aimed to know aggregated IgG-stimulating effects of mesenchymal stem cells in relation to IDO secretion, an immunosuppressant agent. Mesenchymal stem cells was isolated from liposuction waste and characterized based on its surface molecule accrording to ISCT criterias. The confirmed cell culture was used to the next culture by the addition of heat aggragated immunoglobulins gamma (HAGG) for six days. IDO secreted on supernatant culture was collected and assayed by elisa method at 450 nm. The results showed that there was elevated levels of IDO (16.73-49.5%) on culture with HAGG stimulation than without stimulation. This study could be a part of the basicSelama ini sel punca mesenkim telah diketahui mempunyai aktivitas imunosupresan. Secara klinis, sel ini terbukti mampu mampu mengurangi gejala penolakan organ transplan. Pembuktian secara klinis juga sedang dilakukan pada penyakit berbasis inflamasi seperti pada penyakit autoimun yang disebabkan karena kompleks imun, seperti pada lupus eritematosus sistemik (SLE). Penelitian ini bertujuan untuk mengetahui dan mempelajari efek stimulasi sel punca mesenkim dalam hubungannya dengan efek imunosupresan. Sel punca mesenkim diisolasi dari limbah liposuction dan dikarakterisasi berdasarkan molekul permukaannya untuk konfirmasi identitasnya berdasarkan kreiteria ISCT. Selanjutnya sel punca mesenkim dikultur dengan penambahan Heat Aggragated Immunoglobulin Gamma (HAGG) selama 6 hari. Untuk mengetahui sifat imunosupresannya, dilakukan pemeriksaan kandungan senyawa indoleamin 2,3-dioksigenase (IDO) pada supernatan hasil kultur dengan metode ELISA pada panjang gelombang 450 nm. Hasil menunjukkan bahwa terjadi peningkatan kadar IDO pada kultur dengan stimulasi dibanding tanpa stimulasi sebanyak 16,73-49,5 %. Penelitian ini dapat menjadi dasar ilmiah yang memperkuat penggunaan terapi sel punca mesenkim pada penyakit autoimun berbasis kompleks imun

Simple Production Method of Umbilical Cord Derived Mesenchymal Stem Cell Using Xeno-Free Materials for Translational Research

Umbilical cord (UC) is a very promising source of mesenchymal stem cells (MSCs) for allogeneic use, as it can be collected easily without side effects to the donor. In this study, we developed a simple production protocol to get UC-MSCs using xeno-free material to provide safe MSCs for translational research in regenerative medicine. In this study, we used multiple harvest explant method to isolate the MSCs. The medium for isolation and propagation was 10% platelet concentrate (PC) containing alpha MEM. Confluent cultures were harvested, combined and counted. Primary cultures were expanded in T25 flasks (seeding around 5000/cm2) to passage-1 (P-1) and P-2, and the results of P-1 and P-2 cultures were counted. From 5 cm of umbilical cord, we did explant culture in four 24 well plates. The number of harvest per well ranges from 0-5 times i.e. no harvest= 6, once= 20, twice= 34, three times= 27, four times=8, and five times= 1 well(s), respectively. Therefore, we harvested a total of 206 times from the 96 wells, and got a total of 3,595,600 cells from the primary culture. If we use all of the cells from the primary culture for expansion, we will get a total of 47,809,462 cells in passage-1, a total of 1,022,607,122 cells in passage-2. In conclusion, UC-MSCs can be isolated and expanded easily in 10% PC containing alpha MEM, and is suitable to fill the demand of allogeneic MSCs for patient use