TY - JOUR
T1 - Placental chorionic villus explant culture in xeno-free medium
T2 - A preliminary study
AU - Liem, Isabella Kurnia
AU - Adiwinata, Jeanne
AU - Feroniasati, Lauda
AU - Fasha, Iqbal
AU - Sumapradja, Kanadi
PY - 2014
Y1 - 2014
N2 - Aim: to know whether placental chorionic villous-derived stem cells can grow in xenofree medium. In this study, we did explant culture of human chorionic villous in alpha minimal essential medium (MEM) supplemented with human AB serum. Experimental: The chorionic villous was washed briefly, and minced in complete alpha MEM medium, which contained penicillin/streptomycin, amphotericin B, 1% L-glutamine and 10% human AB serum. Two to three explants (diameter 2-3 mm) was placed in each well of a 12 well plate, and several drops of the complete medium were added. Further, the plate was incubated and when the explant attached to the base of the well, 0.5 mL fresh medium was added. Further, medium changes were done every 2-3 days. Observation was done daily to detect cell growth, and cell morphology. The day when fibroblastic cell growth and appearance of various morphologies of cells in each well was first detected was noted and tabulated. Results: Two types of cells were grown, i.e. fibroblastic and non-fibroblastic cells. The fibroblastic cells began to grow and attach on day-3 to day-5. The various morphologies of cells were apparent on day-5 to day-7. Long-term explant culture, more than four weeks, showed that hematopoietic stem cell derived osteoclast-like multinuclear non-fibroblastic cells were prominent and dominated the culture field. Conclusion: Placental chorionic villous explant culture can grow well in 10% human AB serum containing alpha MEM, but when washing is inadequate, the growing cells were dominated by osteoclastlike cells.
AB - Aim: to know whether placental chorionic villous-derived stem cells can grow in xenofree medium. In this study, we did explant culture of human chorionic villous in alpha minimal essential medium (MEM) supplemented with human AB serum. Experimental: The chorionic villous was washed briefly, and minced in complete alpha MEM medium, which contained penicillin/streptomycin, amphotericin B, 1% L-glutamine and 10% human AB serum. Two to three explants (diameter 2-3 mm) was placed in each well of a 12 well plate, and several drops of the complete medium were added. Further, the plate was incubated and when the explant attached to the base of the well, 0.5 mL fresh medium was added. Further, medium changes were done every 2-3 days. Observation was done daily to detect cell growth, and cell morphology. The day when fibroblastic cell growth and appearance of various morphologies of cells in each well was first detected was noted and tabulated. Results: Two types of cells were grown, i.e. fibroblastic and non-fibroblastic cells. The fibroblastic cells began to grow and attach on day-3 to day-5. The various morphologies of cells were apparent on day-5 to day-7. Long-term explant culture, more than four weeks, showed that hematopoietic stem cell derived osteoclast-like multinuclear non-fibroblastic cells were prominent and dominated the culture field. Conclusion: Placental chorionic villous explant culture can grow well in 10% human AB serum containing alpha MEM, but when washing is inadequate, the growing cells were dominated by osteoclastlike cells.
KW - Chorionic villous
KW - Explant culture
KW - Human AB serum
KW - Osteoclast-like cells
KW - Placenta
UR - http://www.scopus.com/inward/record.url?scp=84898628340&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:84898628340
SN - 0974-7435
VL - 9
SP - 280
EP - 284
JO - BioTechnology: An Indian Journal
JF - BioTechnology: An Indian Journal
IS - 7
ER -