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ctt-journal > Shakhpazyan N.1 et al. (Abstract)

Shakhpazyan N.1 et al. (Abstract)

Cellular Therapy and Transplantation (CTT), Vol. 3, No. 12
doi: 10.3205/ctt-2011-No12-abstract39

© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported

Abstract accepted for "5th Raisa Gorbacheva Memorial Meeting Hematopoietic Stem Cell Transplantation in Children and Adults", Saint Petersburg, Russia, September 18–20, 2011

Preliminary Program

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Biological features and quality of human placenta-derived mesenchymal stem cells for clinical use

Nicolay K. Shakhpazyan1, Alexey E. Gomzyakov1, Tatiana A. Astrelina1,2, Mariya V. Yakovleva1, Elena V. Boyakova1,2, Elizaveta V. Kleina1, Yanna A. Kruglova1

1Stem Cell Bank, Moscow, Russia; 2Federal Research Center for Pediatric Hematology, Oncology and Immunology, Moscow, Russia

Correspondence: Tatiana A. Astrelina, Stem Cell Bank, Moscow, Russia, 31, Bakinskaya str., Moscow, 115541, Russia, E-mail: t_astrelina@spam is badmail.ru

Abstract

Aim: To develop optimal conditions for expansion of mesenchymal stem cells (MSC) derived from the placenta and to estimate the quality and biological safety for clinical application.

Methods: Isolation of MSC was carried out of the 20 placentas of healthy donor mothers (1 after cesarean section); mean gestational age was 39.5 weeks. Placenta was obtained after informed consent from the donor, without interference in the birth process. Expansion of MSC was performed by standard methods.

Results: Isolation of placental cells was carried out after grinding and processing of 0.1% collagenase solution pancreas crab for 60 minutes at 37°C. Isolation of cells was optimal at 4–5.5 mg of collagenase per 1 g of placenta with a 0.7–1.2 million cells on 1 g of placenta in the samples obtained. The optimal cell density in the culture flask was 150–210 x 103 per cm2. In the process of expansion MSC 88–319 million cells were obtained after 3–5 passages, which corresponds to 1–2 doses per sample. MSC quality was estimated by bacteriological and viral control, determination of the viability of cells with the Trypan Blue, and markers that specifically identify MSCs (positive for CD13, CD29, CD44, CD73, CD 90, CD105, and CD166 and negative for CD14, CD19, CD34, CD133, CD45, and HLA-DR) by flow cytometry. The biological safety of the MSC was analyzed at 4–5 and 9–10 passages by karyotyping (20–30 metaphase cells for each culture were analyzed). After prolonged cultivation, MSC karyotypes did not change and potentially dangerous genetic abnormalities were not identified.

Conclusions: Thus, optimum conditions for expansion of MSC derived from the placenta were developed. The appropriate MSC quality estimation avoids the use of low-quality and biologically hazardous biomaterial for clinical use.

Keywords:
mesenchymal stem cells, placenta, biological characteristics, quality, safety