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Background: The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation. Objective: To observe the effect of UC-MSCs-CM on the interaction between pro-and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC. Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA. Results: The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001). Conclusion: IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes. Key words: Conditioned medium, BAX, Survivin, GDF9, IGF-1.

Background: Mesenchymal stem cells (MSCs) are known to have immunomodulatory, anti-inflammatory, anti-apoptotic, and angiogenesis effects that are useful for relieving inflammation, recovery, and protection of lung tissues in COVID-19 patients. Secretome, a secretory product of MSCs, has several advantages over MSCs. We conducted a study to investigate secretomes' effectiveness and safety profile as a treatment for severe COVID-19.  Methods: A double-blind, multicenter, randomized, placebo-controlled trial was conducted between February 2021 and July 2021 in three top COVID-19 referral hospitals in the Greater Jakarta area, Indonesia. Eligible subjects (n=40) were randomized in a 1:1 ratio to an intervention group (n=20) and a control group (n=20). The primary outcome of this study was the changes in inflammatory markers and the ratio of inflammatory to anti-inflammatory markers. The secondary outcomes of this study included clinical outcome, laboratory outcome, radiological outcome, RT-PCR result conversion, and safety profile of MSC secretome. Results: Our analysis showed that on the 14th day after placebo administration, IL-6 level in the control group was significantly increased [4.110 (2.403-12.820) at baseline to 13.320 (2.958-33.285) on the 14th day after intervention, p=0.017]. The IL-6/IL-10 ratio in the control group was significantly increased (p=0.036) on the 14th day after placebo administration. We also found that most of the subjects who received placebo had high levels of IL-6 and ferritin (p=0.043) on the seventh day after the intervention. However, we found no significant differences in inflammatory marker levels on the seventh day and 14th day after intervention between both groups. There was no adverse event reported. There were no significant differences in the laboratory outcome, radiology outcome, RT-PCR result conversion, and safety profiles between both groups. Conclusions: MSC secretome can control inflammation in patients with severe COVID-19 and has a good safety profile. MSC secretome is a promising treatment modality for severe COVID-19.

One of the main causes of acute respiratory distress syndrome in coronavirus disease 2019 (COVID‐19) is cytokine storm, although the exact cause is still unknown. Umbilical cord mesenchymal stromal cells (UC‐MSCs) influence proinflammatory T‐helper 2 (Th2) cells to shift to an anti‐inflammatory agent. To investigate efficacy of UC‐MSC administration as adjuvant therapy in critically ill patients with COVID‐19, we conducted a double‐blind, multicentered, randomized controlled trial at four COVID‐19 referral hospitals in Jakarta, Indonesia. This study included 40 randomly allocated critically ill patients with COVID‐19; 20 patients received an intravenous infusion of 1 × 106/kg body weight UC‐MSCs in 100 ml saline (0.9%) solution (SS) and 20 patients received 100 ml 0.9% SS as the control group. All patients received standard therapy. The primary outcome was measured by survival rate and/or length of ventilator usage. The secondary outcome was measured by clinical and laboratory improvement, with serious adverse events. Our study showed the survival rate in the UC‐MSCs group was 2.5 times higher than that in the control group (P = .047), which is 10 patients and 4 patients in the UC‐MSCs and control groups, respectively. In patients with comorbidities, UC‐MSC administration increased the survival rate by 4.5 times compared with controls. The length of stay in the intensive care unit and ventilator usage were not statistically significant, and no adverse events were reported. The application of infusion UC‐MSCs significantly decreased interleukin 6 in the recovered patients (P = .023). Therefore, application of intravenous UC‐MSCs as adjuvant treatment for critically ill patients with COVID‐19 increases the survival rate by modulating the immune system toward an anti‐inflammatory state. Acute respiratory distress syndrome in COVID‐19 patients is caused by a cytokine strom. Umbilical cord mesenchymal stromal cell (UC‐MSC) influence proinflammatory Th2 cells to shift to an anti‐inflammatory agent. An UC‐MSC infusion was given for the experimental group, and normal saline for the control group. Our result showed 2.5 times significantly higher survival rate in the experimental group that achieved by modulating the immune system toward anti‐inflammatory state.

The changing of education activities (offline into online) to reduce coronavirus transmission during COVID-19 pandemi has influence on the learning strategies, which ultimately might impact the achievement of learning objectives. Therefore, we conducted a cross-sectional study using a valid ( p  < 0.01; two-tailed Pearson correlation) and reliable ( r  = 0.878 and r  = 0.849; Cronbach Alpha) online questionnaire to evaluate the perspectives of human anatomy lecturers and undergraduate medical students in Indonesia in implementing the online anatomy learning. We also explored their expectations and preferred learning methods after COVID-19 pandemic. Using purposive sampling, 467 respondents, which consisted of 66 lecturers from 41 universities in six islands (Java, Sumatera, Kalimantan, Sulawesi, Nusa Tenggara and Papua) and 401 students from 19 universities in four islands (Java, Sumatera, Kalimantan, Papua) were recruited. A Chi-square test was used to analyze the differences in categorical variables. The lecturers (74.2%) and students (63.1%) agreed that online learning effectively delivered the course material ( p  = 0.095). They (69.7% lecturers and 57.9% students) also agreed that learning time allocation was sufficient ( p  = 0.079); moreover, lecturers (53%) and students (56.1%) had good interaction ( p  = 0.689). Nevertheless, 56.1% lecturers and 63.3% students had problem during online practical sessions. They had different perspectives about issues during online classes (69.7% vs 36.4%; p  < 0.01), motivation improvement (72.7% vs 37.4%; p  < 0.01), and time management (87.9% vs 58.4%; p  < 0.01). According to the location of the university, especially in the student's side, students in Java had higher proportion in the two aspects, i.e. learning material ( p  < 0.01) and lecturer-student interaction ( p < 0.01), and had lower proportion in the problems during online class ( p = 0.003) and practical sessions ( p = 0.008). Majority of the respondents (62,2% lecturers in Java, 71.4% lecturers outside Java, 79.6% Students in Java, and 76.6% students outside Java) preferred the use of blended learning after the pandemic (new normal era) and expected to continue the cadaveric practical sessions (82.2% lecturers in Java, 81.0% lecturers outside Java, 91.1% students in Java, and 78.3% students outside Java). In conclusion, the study showed that the satisfaction toward online anatomy learning related to the subject’s role (lecturer or student) and the university region in some aspects.

Introduction Treatments for AGA have yet to produce satisfactory outcomes and may cause intolerable side effects. Recent studies have reported that adipose tissue-derived stem cell conditioned media (ADSC-CM) could induce hair growth and regeneration. Objective To investigate the efficacy of ADSC-CM combined with minoxidil for hair regeneration therapy in male AGA. Methods This study lasted for 6 weeks. Subjects were divided into two groups: concentrated and non-concentrated ADSC-CM. Scalp was divided vertically in half before intradermal injection was administered from the frontal region of the scalp toward the vertex with a 30G needle, spaced about 1 cm apart. Treatment side received 2 ml of ADSC-CM; the other side was given 2 ml of NaCl 0.9% as placebo. Patients applied 5% minoxidil twice daily post-injection. Improvements were assessed using photographs and trichoscan every 2 weeks. Results Hair count, hair density, and mean thickness increased significantly on both sides after 6 weeks, while vellus rate decreased proportionally with the increase of terminal rate. No statistically significant differences between treatment groups were found. Minimum side effects were reported, and subjects were satisfied with the results. Conclusion Combination of ADSC-CM and minoxidil could be a potential agent for hair regrowth. Follow-up research with extensive populations, longer duration, and different study design may be required to confirm the exact mechanisms of ADSC-CM on hair growth. Trial registration : Clinicaltrials.gov, NCT05296863. Registered 25 March 2022—Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT05296863 .

BackgroundDespite being a common cause of quality-of-life impairment, there are no efficacious therapies that could prevent the progression of knee osteoarthritis (KOA). We conducted an open-label trial of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) and hyaluronic acid (HA) for treating KOA.MethodsThis open-label study was conducted from July 2015 to December 2018 at Cipto Mangunkusumo Hospital, Jakarta, Indonesia. Patients diagnosed with KOA were injected three times, comprising of 10 × 106 units of hUC-MSCs in 2-ml secretome implantation and 2-ml hyaluronic acid (HA) injection in the first week, followed with 2-ml HA injection twice in the second and third week.ResultsTwenty-nine subjects (57 knees) were recruited. Seventeen (58.6%) subjects were male, and the mean age was 58.3 ± 9.6 years. Thirty-three (57.9%) knees were classified into Kellgren–Lawrence grade I–II KOA (mild OA). hUC-MSCs significantly decreased pain measured by visual analogue scale in severe KOA from initial to 6th month follow-up [5 ± 2.97 to 3.38 ± 2.44 (p = 0.035)]. The International Knee Documentation Committee score significantly increased at 6th month follow-up (53.26 ± 16.66 to 65.49 ± 13.01, p < 0.001, in subjects with grade I–II and 48.84 ± 18.41 to 61.83 ± 18.83, p = 0.008, in subjects with severe KOA). The Western Ontario and McMaster Universities Osteoarthritis decreased significantly in both groups from initial to 6th month follow-up (from 22.55 ± 15.94 to 13.23 ± 10.29, p = 0.003, and from 27.57 ± 15.99 to 17.92 ± 19.1, p = 0.003, in those with mild and severe KOA, respectively).ConclusionshUC-MSCs could be a potentially new regenerative treatment for KOA. The maximum effect of hUC-MSCs was achieved after 6 months of injection.Level of evidenceTherapeutic level II.

To evaluate the efficacy and safety of umbilical cord serum eye drops for dry eyes in ocular Stevens-Johnson Syndrome (SJS). A pre-post test study with umbilical cord serum (UCS) eye drop for ocular SJS patient with moderate to severe dry eyes. Study was conducted at Kirana Cipto Mangunkusumo General Hospital from June 2020 to December 2020. A total of five patients (five eyes) with a diagnosis of SJS more than 6 months, dry eye symptoms, and abnormal tear stability test results were included in the study. Each patient was asked to instill UCS drop into the affected eye six times daily. Evaluation of ocular symptoms with ocular surface disease index (OSDI) questionnaires, non-invasive tear break-up time (NIBUT), Schirmer I, and keratoepitheliopathy scores was administered before applying UCS drop and at week 2 and 4 of eye drop use. From June 2020 to December 2020, five eyes of five patients were evaluated in this study. Patients were aged from 22 to 71 years old with history of SJS over periods from 1 to 35 years. Three patients underwent ocular surgeries prior to the study. After four weeks of treatment, symptoms score, Schirmer I, and keratoepitheliopathy scores improved significantly, while NIBUT scores improved insignificantly. No side effects were noted during treatment. Administration of UCS eye drop was effective in improving symptoms and signs of dry eye in chronic SJS patients.

IntroductionThe current ‘gold-standard’ treatment of critical-sized bone defects (CSBDs) is autografts; however, they have drawbacks including lack of massive bone source donor site morbidity, incomplete remodeling, and the risk of infection. One potential treatment for treating CSBDs is bone marrow-derived mesenchymal stem cells (BM-MSCs). Previously, there were no studies regarding the use of human umbilical cord-mesenchymal stem cells (hUC-MSCs) for treating BDs. We aim to investigate the use of allogeneic hUC-MSCs for treating CSBDs.MethodWe included subjects who were diagnosed with non-union fracture with CSBDs who agreed to undergo hUC-MSCs implantation. All patients were given allogeneic hUC-MSCs. All MSCs were obtained and cultured using the multiple-harvest explant method. Subjects were evaluated functionally using the Lower Extremity Functional Scale (LEFS) and radiologically by volume defect reduction.ResultA total of seven (3 male, 4 female) subjects were recruited for this study. The subjects age ranged from 14 to 62 years. All seven subjects had increased LEFS during the end of the follow-up period, indicating improved functional ability. The follow-up period ranged from 12 to 36 months. One subject had wound dehiscence and infection, and two subjects developed partial union.ConclusionUmbilical cord mesenchymal stem cells are a potential new treatment for CSBDs. Additional studies with larger samples and control groups are required to further investigate the safety and efficacy of umbilical cord-derived mesenchymal stem cells for treating CSBDs.

To compare the viability and characteristics of umbilical cord mesenchymal stromal cells (UC-MSCs) when they were kept in culture in three types of αMEM after confluence for various time points. Viability and characteristic assessments were done after keeping UC-MSC confluent cultures for 0, 24, 48, 72, and 96hours in αMEM with glutamax (Himedia), and without glutamax (Himedia or Gibco).There were thirty cultures, which each ten of them were kept in the three types of basal media, and for each time point and type of basal medium, two flasks were harvested. All viability and characteristic assessments were done after harvest. Viability assessments were done in duplicate, while characteristics assessment by flowcytometry for CD73, CD90 and CD105 were done only once.Further, differences between the various time points in terms of viability and CD73, CD90 and CD105 percentage after various time points were compared and tested by appropriate statistical analysis. Viability was >70% upto 72 hours and 48 hours when the cells were kept in Himediaglutamax containing αMEM and αMEM without glutamax respectively. Flow cytometry showed that CD73, CD90 and CD negative percentage did not differ to initial percentage, but after 24 hours CD105 was slightly decreased to > 60% in Himediaglutamax containing αMEM, while in the two αMEM media without glutamaxthe percentages were below 40%. For our UC-MSC culture, glutamax containing αMEM is better compared to αMEM without glutamax as temporary storage solution, and storage should be restricted to 24 hours.

Background: Lipoaspirate contains noxious substances derived from liposuction. Therefore, extensive washing is recommended before the lipoaspirate is processed further for culture or fat grafting. Washing a small amount of lipoaspirate may not pose a problem, but washing a large volume of lipoaspirate may be cumbersome, time consuming, and requires a lot of phosphate buffered saline (PBS). Objective: To introduce a simple method for lipoaspirate washing using fine-mesh stainless-steel tea or coffee filter, a small tea spoon, and a porcelain bowl. Methods: The filter was used to collect the adipose tissue fragments. Further washing of the fragments was achieved by soaking the adipose tissue containing filter in a PBS containing porcelain bowl and stirring using a small tea spoon to transfer the contaminating materials to the PBS. Enzymatic processing to dissociate the cells from the tissue and primary cultures was conducted as usual in MesenCult. Results: Using the equipment mentioned above, the adipose tissue fragments were readily separated from the blood, free lipids, anesthetics, and other noxious material in the liquid portion. This simple method saves time and PBS compared with previously described methods. Further enzymatic processing produced sufficient cells to be cultured, and culture results showed plastic adherent cells on day 2 that became confluent on day 6. Conclusion: Lipoaspirate washing using a fine mesh stainless steel filter is time saving and produced cells that grow well in MesenCult.