Systems approach to the guarantee of quality of mesenchymal stem cells of bone marrow for the clinical use

Mesenchymal stromal/stem cells (MSCs) of bone marrow
(BM) origin not only provide the supportive microenvironmental
niche for hematopoietic stem cells (HSCs) but are also capable
of differentiating into various cell types of mesenchymal
origin, such as bone, fat, and cartilage. The role is known
for bone marrow-derived MSCs in reducing the incidence and
severity of graft-versus host disease (GVHD) during allogeneic
transplantation. Purpose: to estimate quality and biological
safety the MSC of bone marrow for the transplantation.
Were analyzed data expansion MSC of 94 BM at the Stem Cell
Bank of Moscow between 2007 and 2010 on the registered
medical technologies. MSC were revealed in the native form
or frozen in liquid nitrogen. Quality, MSC was evaluated with
of the bacteriological and virusological control; determined
the viability of cells with the trypan blue and 7AAD; markers
that specifically identifies MSCs: CD73+; CD 90+; CD105+;
CD45-; CD34-; CD14; CD133-; CD19-; HLA DR- by flow
cytometry. The biological safety (karyotype) was analyzed by
G-banding technique; 15-30 metaphase cells for each culture
were analyzed. To analyze the level aneuploidy, fluorescent in
situ hybridization (FISH) with chromosome enumeration probes
(CEP) studies was performed. Are developed the documents,
which regulate the stages of the work of expansion MSC of
BM. Were prepared 71 MSC of BM (157 doses) and were
used for allogeneic transplantation 23 to patients (70
doses MSC) for the purpose of adherence HSC, reducing
the incidence and severity of GVHD. In the majority of the
cases of transplantation MSC it was carried out at the
dose of 2 ×106/kg. There were no acute reactions during the
transplantation MSC BM. Thus, the estimation of quality and
safety MSC of BM for allogeneic transplantation, included: a
conducting of documentation on GMP to standards, inspection
of a quantity of cells for achievement of optimum dose,
bacteriological and virusological control, confirmation the
markers that specifically identifies MSCs and the estimation
of biological safety.