Publication Detail

Journal/Magazine : International Journal of Pharm Tech Research; 2014 (August-Sept); 6 No.4; 1202-1208
Umbilical cord derived stem cell culture: Multiple-Harvest explant method
Author : Jeanne Adiwinata Pawitan, Isabella Kurnia Liem, Ecie Budiyanti, Iqbal Fasha, Lauda Feroniasanti, Taufik Jamaan, Kanadi Sumapradja
Departemen :Histology, Level : International, Article Category : research
Medical Writer : yes, Research Type : biomedic

Abstract: 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.

Key words: umbilical cord, explant culture, multiple harvests, stem cell, thrombocyte concentrate


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