Cipto Mangunkusumo Central Hospital

Jakarta, Indonesia

Cipto Mangunkusumo Central Hospital

Jakarta, Indonesia
SEARCH FILTERS
Time filter
Source Type

Budiyanti E.,Jl. Salemba 6 | Liem I.K.,University of Indonesia | Pawitan J.A.,University of Indonesia | Wulandari D.,University of Indonesia | And 3 more authors.
International Journal of Research in Pharmaceutical Sciences | Year: 2015

Previous studies used fetal bovine serum (FBS) containing medium for umbilical cord derived mesenchymal stem cell culture. Xenoproteins in FBS may be incorporated into the cultured cells and cause immune rejection, when the cells are used in patients. Therefore, the aim of this study was to compare propagation performance of umbilical cord derived stem cells that were cultured in various platelet rich plasma (PRP) containing media and FBS derivate containing commercial medium.This study was an in vitro analytical study on multiple harvest explant culture derived stem cells, which compared the stem cell population doubling time (PDT) of passage 1-3 cultures in human AB PRP containing media (low glucose and high glucose DMEM, and αMEM) compared to xenopretein containing commercial medium, MesenCult®. Overall, the lowest PDT was achieved by αMEM-PRP, and PDT in DMEM LG-PRP and αMEM PRP was comparable to those of MesenCult®. Features of cluster of differentiation (CD) expressions in P-1 suggest that the umbilical cord derived stem cells obtained from multiple harvest explant cul-ture were not homogenous for MSCs. However, MSCs tended to become more homogenous with passages, espe-cially for cells that were cultured in MesenCult®, followed by those cultured in αMEM-PRP. In conclusion, the best propagation performance of umbilical cord derived stem cells passage 1-3 cultures was achieved in PRP containing αMEM. © JK Welfare & Pharmascope Foundation.


Pawitan J.A.,Jl. Salemba | Pawitan J.A.,Cipto Mangunkusumo Central Hospital | Liem I.K.,Cipto Mangunkusumo Central Hospital | Budiyanti E.,Jl. Salemba 6 | And 4 more authors.
International Journal of PharmTech Research | Year: 2014

Published methods of umbilical cord explant culture require removing the explant after the MSCs are confluent. However, we observed that the explants were still attached after the MSCs were detached using TrypLE Select. Therefore, the objective of this study was to do explant cultures in xeno-free media, and reculture the explants after the first harvest to test whether the explant could grow further to yield more stem cells. In this study, umbilical cord explant culture were done in 10% platelet rich plasma (PRP) containing alpha minimal essential medium (αMEM) or Dulbecco's modified Eagle's medium (DMEM), or 10% human AB or cord blood serum (CBS) containing αMEM. Three explants were cultured in each well of a twelve well plate in triplicate. The cells were harvested using TrypLE Select, and cell yield was counted after cell growth achieved 80-90% confluence. The remaining attached explants were re-cultured in the respective media for several times after harvesting, until the explants were detached or until there was no more cell growth from the explants. Cell counts for every harvest were noted. Cumulative cell yield from each well for each medium was computed and tabulated. Our results showed that the time needed for cell growth was variable between explants. Therefore, in one well, the three explants might grow with different speed, and in many instances, harvests were done before optimal growth in all explants occurred. The maximal number of harvest was four times, which occurred in 10% human AB serum supplemented αMEM (two times) followed by two harvest in 10% autologous CBS supplemented αMEM, and yielded a cumulative amount of 144,000 viable cells in 37 days. In conclusion, we have developed a multi-harvest explant method to culture umbilical cord stem cells in various xeno-free media that yield far more numerous viable cells compared to standard explant method.

Loading Cipto Mangunkusumo Central Hospital collaborators
Loading Cipto Mangunkusumo Central Hospital collaborators