High transduction efficiency of circulating first trimester fetal mesenchymal stem cells: potential targets for in utero ex vivo gene therapy

Abstract

We recently reported the existence of fetal mesenchymal stem cells in first trimester fetal blood. Here we demonstrate that fetal mesenchymal stem cells from as early as eight weeks of gestation can be retrovirally transduced with 99% efficiency without selection. Circulating fetal mesenchymal stem cells are known to readily expand and differentiate into multiple tissue types both in vitro and in vivo, and might be suitable vehicles for prenatal gene delivery. With advances in early fetal blood sampling techniques, we suggest that genetic disorders causing irreversible damage before birth could be treated in utero in the late first/early second trimester by genetically manipulated autologous fetal stem cells.

Direct Muscle Delivery of GDNF With Human Mesenchymal Stem Cells Improves Motor Neuron Survival and Function in a Rat Model of Familial ALS

“…hMSC were isolated from neonatal bone marrow aspirates from healthy donors after informed consent and confirmed by immunophenotyping and multilineage differentiation [27-29]. This line was then genetically modified to express green fluorescent protein (GFP) using retrovirus (hMSC GFP ) [27].…”

“…This line was then genetically modified to express green fluorescent protein (GFP) using retrovirus (hMSC GFP ) [27]. hMSC GFP were infected with a lentiviral construct encoding GDNF under the control of phosphoglycerol kinase (pgk) promoter (Fig.…”

“…Human mesenchymal cells were established using methods previously described [27-29]. These cells were derived from neonatal bone marrow aspirates from healthy donors after informed consent.…”

“…When colony size exceeded >500 cells, single colonies were trypsinized and collected using clonal cylinders. The mesenchymal nature of adherent nucleated cells was confirmed by immunophenotyping and multilineage differentiation [27-29]. In the current study, we used a hMSC line genetically modified to express green fluorescent protein (GFP) using retrovirus (N97-GFP; hMSC GFP ) [27].…”

Concise Review: Multipotent Mesenchymal Stromal Cells in Blood

“…Besides the BM and PB, other organs and tissues in adults were also shown to be sources of MSCs, including pleural cavity, spleen, thymus, peritoneal cavity, lymph node, adipose tissue, muscle, brain, and exfoliated deciduous teeth [1, 8, 57, 86, 142, , –145]. Although Wexler et al [73] failed to identify MSCs in umbilical cord blood (UCB) from full‐term deliveries, the isolation of MSCs from UCB was successful in other laboratories [62, 146, , –149]. It was found that human first‐trimester fetal blood contained more MSCs than blood from the second and third trimesters [147].…”