Jeanne Adiwinata Pawitan

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.

Platelet Rich Plasma in Xeno-Free Stem Cell Culture: The Impact of Platelet Count and Processing Method

BACKGROUND Stem cell culture for regenerative medicine needs platelet rich plasma (PRP) as fetal bovine/calf serum (FBS/FCS) substitute. However, the various studies used various protocols in preparing and processing the PRP. This study aimed to compare and conclude the most effective and efficient protocol. METHODS we searched in vitro studies that used human PRP as FBS/FCS substitute to culture human cells, and compared the various available protocols to identify the easiest and effective protocols for the preparation of PRP and the release of the growth factors (GFs) to support the highest cell growth in stem cell culture. RESULTS ten studies fulfilled the selection criteria and were included in the analysis. DISCUSSION Almost all studies on bone marrow mesenchymal stem cell (BM-MSC) and adipose stem cell (AT-SC) showed that platelet lysate and/or activated platelet releasate were superior or at least the same as either FBS or FCS, except for one study that got different results on human AT-SC. Several studies showed that either 5% activated PRP (aPRP) or platelet lysate (PL) was sufficient to support cell growth, or even better when they were compared to 10% FBS, while higher concentrations were counterproductive. However, some studies showed that 10% aPRP or PL was needed. The difference between studies was due to the difference in either the PRP preparation from blood and in the PRP processing to release the GFs, which yield various GF concentrations. CONCLUSION In conclusion, studies are needed to reveal the optimal final platelet counts for the various PRP processing methods for various kinds of cells. The easiest PRP processing is freezing to -20°C followed by thawing, or thrombin activation using a final concentration of 100U/mL.

Prospect of Adipose Tissue Derived Mesenchymal Stem Cells in Regenerative Medicine

Adipose tissue derived mesenchymal stem cells (AT-MSCs) have similar properties to the previously characterized bone marrow mesenchymal stem cells (BM-MSCs). However, adipose tissue is easier to get in larger volumes, the procedure is less painful and at lower risks, and yield more stem cells compared to bone marrow. Moreover, considering the various differentiation and angiogenic potentials, the ease in collecting the samples, and the immuno-suppressive properties, adipose tissue stem cells are very promising for regenerative medicine, and therefore will be the preferred source of MSCs for future clinical use.

Prospect of cell therapy for Parkinson's disease

The hallmark of Parkinsons disease is on-going degeneration of dopaminergic neurons in the substantia nigra, which may be due to various etiologies. Various approaches to alleviate symptoms are available, such as life-long pharmacological intervention, deep brain stimulation, and transplantation of dopaminergic neuron-containing fetal tissue. However, each of these approaches has a disadvantage. Several studies have shown that various kinds of stem cells, induced pluripotent stem cells, and other cells can differentiate into dopaminergic neurons and may be promising for treating Parkinsons disease in the future. Therefore, this review addresses those cells in terms of their prospects in cell therapy for Parkinsons disease. In addition, the need for safety and efficacy studies, various cell delivery modes and sites, and possible side effects will be discussed.

The possible use of RNA interference in diagnosis and treatment of various diseases

Aims:  To address the possible use of RNA interference in diagnosis, prognosis and therapy of various diseases and explain the obstacles in RNA interference based therapy.

Modified Masquelet technique using allogeneic umbilical cord-derived mesenchymal stem cells for infected non-union femoral shaft fracture with a 12 cm bone defect: A case report

Highlights • A case of infected non-union fracture with critical-sized bone defect.• Modified Masquelet technique was used to overcome the defect.• The addition of allogenic UC-MSCs, BMP-2 and hydroxyapatite encourages healing.• This technique may serve as an effective way to overcome large defects.

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 patients own cell can be developed. However, iPSC from the patientss 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.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 patients own cell can be developed. However, iPSC from the patientss 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.

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.

Bone Marrow Stem Cells Anti-liver Fibrosis Potency: Inhibition of Hepatic Stellate Cells Activity and Extracellular Matrix Deposition.

Transplantation of bone marrow derived stem cells (BMSCs) has been reported inhibits liver fibrosis. Several in vitro studies by co-culturing BMSCs and hepatic stellate cells (HSCs) indirectly or directly in 2D models showed inhibition of HSC as the key player in liver fibrosis. In this study, we investigated direct effect of BMSCs on HSCs by co-culturing BMSCs and HSCs in 3D model as it represents the liver microenvironment with intricate cell-cell and cell-matrix interactions. Primary isolated rat HSCs and BMSCs were directly co-cultured at 1:1 ratio with hanging drop method. The monoculture of rat HSCs served as positive control. Mono-culture and co-culture samples were harvested on day 3, 5 and 7 for histological analysis. The samples were analyzed for extracellular matrix deposition by Masson’s Trichrome staining, tenascin-C immunocytochemistry, resting HSC’s state as shown by positive Oil Red O stained cells. Our results indicated CD90+CD34− BMSCs anti-liver fibrosis potency as evidenced by higher proportion of Oil Red O-positive cells in the co-culture group compared to the monoculture group and the significant decrease in extracellular matrix deposition as well as the decrease in tenascin-C expression in the co-culture group (p<0.05) compared to the monoculture group. These findings demonstrate that BMSCs have a potential therapeutic effect against liver fibrotic process through their capacity to inhibit HSCs activation and their effect in minimizing extracellular matrix deposition.

Various stem cells in acupuncture meridians and points and their putative roles

Traditional Chinese and Korean medicine uses various manipulations on acupuncture points/acupoints that are located along imaginary lines on the surface of a human body, which are called ‘meridians’. Acupuncture has been used from the ancient time till now to cure various diseases, including for the purpose of regenerative medicine. In various studies, meridians are alternatively called as Bong-Han ducts, primo vessels, or hyaluronic-acid rich ducts, while acupoints are called Bong-Han corpuscles, primo nodes, or hyaluronic-acid rich nodes. Meridians and acupuncture points form a system that is now called primo vascular system (PVS), which is claimed to contain various kinds of stem cells. The stem cell size is between 1-5 microns. The smallest is the primo microcells that have a putative role in regeneration. Other stem cells are adult pluripotent and hematopoietic stem cells that play a role in extra bone marrow hematopoiesis. The presence of PVS has been reproduced by many studies. However, the various stem cells need further studies to prove their existence and function, and harvesting PVS to isolate the stem cells might harm the health of the donor.