Optimization of conditions of skin and gingival mucosa derived human fibroblasts obtainment and cultivation

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Abstract

For optimization of conditions of skin and gingival mucosa derived human fibroblasts obtainment and cultivation in this work we performed comparative analysis of 11 fetal bovine serums (from different manufacturers) and autologous human serum. Our analysis included also comparison of 3 serum-free media and 4 serum-reduced media. Evaluation criterion at the step of primary cell culture was colony-formation efficiency and during subsequent cultivation - fibroblasts population doubling time. Results revealed that ability of skin-derived fibroblasts to form colonies could vary 4 and more times and gingival mucosa derived fibroblasts - 1,5-1,8 times depending on used for cultivation type of serum or medium. Clonal analysis showed that fibroblasts in culture form 3 types of colonies consisting of different amount of cells and different phenotype. Ratio of colonies could be variable and depend on type of used medium. It was noticed during cultivation that addition of different sera to standard medium (DMEM) led to changing of cell's doubling time. In general, successful obtainment of fibroblasts primary culture determined by selection of appropriate serum and medium providing development of large colonies of rapidly proliferating cells. It was noticed during cultivation that addition of different sera to standard medium (DMEM) led to changes in cell's doubling time.

About the authors

V. L Zorin

Human Stem Cells Institute, Moscow, Russia; A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

P. B Kopnin

Human Stem Cells Institute, Moscow, Russia; N.N. Blokhin Cancer Research Center of RAMS, Moscow, Russia

A. I Zorina

Human Stem Cells Institute, Moscow, Russia

I. I Eremin

Human Stem Cells Institute, Moscow, Russia; A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

N. L Lazareva

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

T. S Chauzova

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

D. P Samchuk

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

A. P Petrikina

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

P. S Eremin

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

I. N Korsakov

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

O. S Grinakovskaya

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

E. V Solovieva

Human Stem Cells Institute, Moscow, Russia

K. V Kotenko

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

A. A Pulin

A.I. Burnazyan Federal Medical Biophysical Center FMBA of Russia, Moscow, Russia

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