A Novel Synthetic, Xeno‐Free Biomimetic Surface for Serum‐Free Expansion of Human Mesenchymal Stromal Cells

Abstract

diseases for which no therapies currently exist including graft-versus-host disease, [1,2] autoimmune diseases, [3] liver diseases, [4] orthopedic injuries, [5] cardiovascular diseases, [6] and cancer. [7] But despite high interest and a large number of clinical trials, only three hMSC-based therapies have been clinically approved yet. This might be largely due to the fact that their medical application still faces many challenges including heterogeneity in therapeutic targets, routes of delivery, range of doses, and manufacturing protocols. [8] With minimal effective doses ranging from 70 to 900 million cells per patient extensive ex vivo expansion and large-scale production are required. [9] Furthermore, the therapeutic efficacy and proliferation capacity of hMSCs decline during aging and passaging, and thus optimal growth conditions in vitro are urgently needed to produce adequate numbers of high-quality cells for clinical use. [10,11] hMSCs have been isolated from a wide range of adult and perinatal tissues and are advantageous to induced pluripotent and embryonic stem cells by not bearing the risk of tumorigenesis and their easy applicability. [12,13] They are a heterogeneous population of tissue-specific stem/progenitor cells that possess the ability to recreate the tissue from which they are isolated. [14] The self-renewal and differentiation potential of hMSCs are influenced by not only their tissue of origin but also donor age, genetics, exposure to environmental stress, and passaging in vitro. [15,16] Since their morphology can easily be mistaken for fibroblasts, the International Society for Cellular Therapy defined minimal criteria for hMSCs in vitro: 1) adherent growth in plastic culture dishes, 2) differentiation capacity into adipocytes, osteocytes, and chondrocytes, and 3) expression of CD73, CD105, and CD90 in at least 95% of the cell population and absence of the hematopoietic markers CD45, CD34, CD11b, or CD14. [17] hMSCs express also other cell surface molecules, for example, CD146, intercellular adhesion molecule (ICAM), CD271, Nestin, and leptin receptor that define them as multipotent tissue-specific stem/progenitor cells. The most frequently used source of hMSCs for clinical trials is bone marrow, from which cells can be isolated using Ficoll density gradient centrifugation. The subsequent ex vivo expansion is facilitated by nutrients, growth factors, and extracellular matrix (ECM) proteins present in the undefined soluble component of Human mesenchymal stromal cells (hMSCs) have enormous potential for the treatment of various inflammatory and degenerative diseases. Their manufacturing for cell-based therapies requires extensive ex vivo expansion and optimal growth conditions. To support cell adhesion, spreading, and growth in serum-free culture conditions, the applied plasticware needs to be functionalized with essential biochemical cues. By employing a recently developed screening tool, a chemically defined functional matrix composed of dextran sulfate and a bone-related extracellular ma...

A chemically defined biomimetic surface for enhanced isolation efficiency of high-quality human mesenchymal stromal cells under xeno-/serum-free conditions

“…We and others have previously reported that BM-MSCs expanded in serum-free medium show a smaller cell size than MSCs expanded in serum-containing medium 20,21 . Similarly, cells isolated using the isoMATRIX in combination with xeno-/serum-free medium were significantly smaller (16 ± 0.8 µm) compared to those isolated under serum-containing conditions (20 ± 0.8 µm; Fig.…”

“…We previously developed myMATRIX MSC, a surface coating optimized for MSC expansion, which showed suboptimal support during MSC isolation from bone marrow aspirates (data not shown). To identify a new biomatrix that can aid MSC isolation, we selected 12 candidates, previously identified from our in-house screening tool (screenMATRIX), that supported efficient MSC expansion 20 . In preliminary isolation tests, the use of a biomatrix composed of dextran sulfate and a fibronectin-derived peptide-conjugate resulted in the best cell morphology and highest isolated cell number (data not shown).…”

“…This matrix is formed by sulfated glycosaminoglycan (GAG) interactions with a fourarm polyethylene glycol-modified with a biofunctional peptide 19 . It promotes cell adhesion and expansion using different media compositions and supports the longterm culture of MSCs in a serum-free medium with robust growth and high viability while maintaining their differentiation and immunomodulatory capacity, characteristic cell morphology, and the expression of key stemness markers 20 .…”

Screening Arrays of Laminin Peptides on Modified Cellulose for Promotion of Adhesion of Primary Endothelial and Neural Precursor Cells

“…[ 5,6 ] Alternatively, synthetic materials can also be employed. [ 7–12 ] While proteins or ECM extracts can enable effective cell adhesion and most faithfully mimic the cellular microenvironments, synthetic materials offer more defined substrates with low batch‐to‐batch variation facilitating their applications especially for clinical purposes. [ 13,14 ]…”

“…Many primary and stem cells are anchorage‐dependent cell types and require a certain degree of adhesion to a solid culture support to ensure cell survival, growth, and development in vitro. [ 10,15–17 ] To enable cell adhesion, synthetic materials can be modified with peptides ligands, which are specific adhesion sequences derived from ECM proteins. The first reported and most frequently used sequence Arg‐Gly‐Asp (RGD) has been identified by Ruoslahti and colleagues in 1984.…”