Comparison of fetal bovine serum and platelet-rich plasma on human lipoaspirate-derived mesenchymal stem cell proliferation.

Background: Lipoaspirate-derived stem cells (LSCs) are very promising for regenerative medicine, e.g. to treat acute myocard infarction. Fetal bovine serum (FBS) is commonly used to propagate the LSCs. However, for its clinical application, FBS contains xeno-proteins that are potential to elicit immune rejection in patients. Platelet rich plasma (PRP) is one of the candidates to replace FBS. This study was aimed to compare the proliferation of LSCs cultured in 5% PRP, 10% PRP, and FBS containing medium (MesenCult®). Methods: LSCs were cultured in 5% PRP/DMEM, 10% PRP/DMEM, and MesenCult®. After the primary culture reached its confluency, cells were harvested using TrypLE Select and seeded (around 20,000 viable cells) in new vessels in the same media. Passages were done until passage-5, with six replications. Population doubling time (PDT) of the three groups were analyzed using Kruskal Wallis test. Results: LSCs showed different proliferation rates when cultured in 5% PRP/DMEM, 10% PRP/DMEM, and MesenCult®. PDT of the three experimental groups in passage 1-5 were significanly different (p < 0.05), with the lowest rank was cultured in medium of 10% PRP/DMEM. Conclusion: The results suggest that 10% PRP/DMEM can be used as an alternative to replace FBS in LSC culture.

Effect of adipose tissue processing procedures in culture result: a study preliminary.

Background: There are various methods of processing adipose tissue before culture, depending on the adipose tissue samples. The aim of this study is to compare several modifications of culturing and sub-culturing procedures of adipose tissue to fit the condition in our laboratory. Method: This is a descriptive study that was done in the Immunology and Endocrinology Integrated Laboratory, University of Indonesia, from October 2009 to April 2010. Three adipose tissue processing procedures, various amount of seeding and two subculture methods were compared in term of cell yield and time needed. In the first procedure, collagenase-1 digestion was done in 30minutes, cell seeding were 24,000 and 36,000 per flask; in the second procedure, collagenase-1 digestion was done in 60minutes, cell seeding were 24,000, 48,000, and 72,000 per flask; and in the third procedure, the adipose tissue remnants from the first procedure were again digested for another 45 minutes, cell seeding were 74,000, and 148,000 per flask. Difference in subculture methods were the presence or absence of washing step. Result: Procedure 1 yielded the lowest amount of cell, and after culture, the cells grew very slow, and was contaminated before harvest of primary culture. Procedure-2 and -3 succeeded to yield primary cultures. Some of the cultures were contaminated, so that further subculture was not applicable, and only one tissue processing procedure (procedure 2: 60 minute collagenase-1 digestion, without lysis buffer, cell seeding 48,000 and 72,000) could complete the three subcultures. Though some of the procedures could not be completed, final result could be concluded. Conclusion: In this preliminary study, 60 minute colagenase-1 digestion with intermittent shaking every 5 minutes and cell seeding around 50,000 or more, followed by subculture method without washing step gave the best result.

Cytology technique: development of a simple spot method for cultured cell suspension.

Aim To develop a simple spot method to attach cultured cells in suspension on to a glass slide. Methods We compared three approaches using both conventional and special glass slide (Shandon-Polysin)., either without additional fetal bovine serum (FBS), or with addition of 3 or 10 μl of FBS to a 20 μl sample (altogether there were six approaches). The slides were examined qualitatively for the background color, boundary color and intactness, and whether there were folded and detached parts. Further, for each slide, the attached intact cells were counted, and the percentage of attached intact cells per number of spotted cells was calculated. The difference in attach intact cells between different approaches was analyzed by ANOVA using SPSS 13.0 for windows. Results There were no significant difference in the percentage of attached intact cells between the six approaches (P= 0.804), though the approach using special glass slide without additional FBS (FBS final concentration 5%) yield the highest percentage of attached intact cells, showed clean background without folded parts. Conclusions We have developed a simple spot method for cultured cell suspension, and the best approach to make spot specimen is using special glass slide with 5% FBS in the cell suspension.

Association between presence in lectures and student knowledge gain in problem based learning: experience in Neuropsychiatry module, FMUI, International Program.

Aim This study was conducted to determine the association between presence in lecture and examination scores. Methods This was a cross sectional study, conducted in the Faculty of Medicine University of Indonesia, International Class Program, from November 2007 to May 2008. The subject’s inclusion criterion was FMUI International class students enrolled in the Neuropsychiatry module in 2007/2008. we excluded students who did not attend the examinations i.e. multiple choice questions (MCQ)-1, -2, or essay. The data collected were presence in lecture and examination scores. Data analysis was done using SPSS linear regression to see the association between presence in lectures and exam scores. Results The results showed that the R and P of the association between presence in lectures and exam scores were (R= 0.121, P= 0.413), (R= 0.212, P= 0.148), (R= 0.260, P= 0.075), and (R= 0.280, P= 0.054) for MCQ1, MCQ2, essay, and mean exam scores respectively. Conclusion We failed to show the association between presence in lectures and student knowledge gain. This result might be due to the many factors discussed that were not analyzed in this study.

Language barrier, funding, communication and time management problems found in the Medical Journal of Indonesia, 1995 - 2001.

The objective of this study is to assess the problems encountered in the management of Medical Journal of Indonesia and whether any progress has been achieved during the past 7 years. A retrospective descriptive study was done in the MJI. Available data used as parameters were: number of articles submitted and author's home institution, number of articles submitted and rejected, cause/criteria of rejection, qualification of reviewers and editors, duration of manuscript processing, and whether pre-scheduled time is met in the Med J Indones. For this purpose, all manuscripts from January 1995- January 2001 were evaluated. In addition, recent problems encountered were noted. Problems encountered were limited supply of manuscript that might be due to language barrier, decrease in circulation due to limited budget, communication problems due to seniority, and increase in average processing time that might be due to restriction in the time allocated for MJI Number of articles submitted by authors originating from national versus foreign institutions in 1995-1997 was 101 versus 16, and in 1998 - 2000 were 124 versus 84. Articles rejected were ±26.4 % (1999-2001) and the most frequent causes of rejection were insignificancy of substance and improper data analysis. As for the qualification of editors there was an increase in the number of PhDs and professors. Average processing time is approximately 1.5 times the scheduled time. It seems that the 4 main problems encounteredwere (l.l Language barrier, (2) Funding, (3) Communication problems and (4) Time management; and though not impressive, there are definitely positive changes in the number of manuscipts submitted, qualification of editors and peer reviewers. Average processirtg time is about the same, and the prescheduled time of editing is yet to be met.