Microcarrier choice and bead-to-bead transfer for human mesenchymal stem cells in serum-containing and chemically defined media

Leber, Jasmin; Barekzai, Jan; Blumenstock, Miriam; Pospisil, Björn; Salzig, Denise; Czermak, Peter

doi:10.1016/j.procbio.2017.03.017

Cited by 57 publications

(57 citation statements)

References 37 publications

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“…However, it was reported that cell attachment and growth on microcarriers was slower on using CDM in comparison to FBS‐based setup. This was attributed to the lack of various growth factors which are integral within a serum‐based medium . We demonstrated that replacing FBS with HPL as culture medium for spinner flask‐based expansion of hPDCs on collagen microcarriers had multiple positive effects.…”

Section: Resultsmentioning

confidence: 99%

Human Platelet Lysate Improves Bone Forming Potential of Human Progenitor Cells Expanded in Microcarrier-Based Dynamic Culture

Gupta

Hall

Geris

et al. 2019

Stem Cells Translational Medicine

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Xenogeneic‐free media are required for translating advanced therapeutic medicinal products to the clinics. In addition, process efficiency is crucial for ensuring cost efficiency, especially when considering large‐scale production of mesenchymal stem cells (MSCs). Human platelet lysate (HPL) has been increasingly adopted as an alternative for fetal bovine serum (FBS) for MSCs. However, its therapeutic and regenerative potential in vivo is largely unexplored. Herein, we compare the effects of FBS and HPL supplementation for a scalable, microcarrier‐based dynamic expansion of human periosteum‐derived cells (hPDCs) while assessing their bone forming capacity by subcutaneous implantation in small animal model. We observed that HPL resulted in faster cell proliferation with a total fold increase of 5.2 ± 0.61 in comparison to 2.7 ± 02.22‐fold in FBS. Cell viability and trilineage differentiation capability were maintained by HPL, although a suppression of adipogenic differentiation potential was observed. Differences in mRNA expression profiles were also observed between the two on several markers. When implanted, we observed a significant difference between the bone forming capacity of cells expanded in FBS and HPL, with HPL supplementation resulting in almost three times more mineralized tissue within calcium phosphate scaffolds. FBS‐expanded cells resulted in a fibrous tissue structure, whereas HPL resulted in mineralized tissue formation, which can be classified as newly formed bone, verified by μCT and histological analysis. We also observed the presence of blood vessels in our explants. In conclusion, we suggest that replacing FBS with HPL in bioreactor‐based expansion of hPDCs is an optimal solution that increases expansion efficiency along with promoting bone forming capacity of these cells. Stem Cells Translational Medicine 2019;8:810&821

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Section: Resultsmentioning

confidence: 99%

Human Platelet Lysate Improves Bone Forming Potential of Human Progenitor Cells Expanded in Microcarrier-Based Dynamic Culture

Gupta

Hall

Geris

et al. 2019

Stem Cells Translational Medicine

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“…Lastly, a significant advantage of MC based cultures is that the growth surface provided to the cells can be increased by simply adding new MCs to the culture, as it has been established that cells are able to migrate from bead to bead and populate newly added microcarriers (35)(36)(37)(38)(39). This phenomenon, commonly referred to as "bead to bead transfer, " can be explained by two main mechanisms: cells detaching from a confluent MC and reattaching onto other MCs, or cells forming bridges between MCs upon collision (40). It has been shown that successively transferring a small proportion of near-confluent MCs (10-25%) into a new vessel loaded with fresh MCs leads to a decrease of lag phase and an increase of the overall yield (35,38).…”

Section: Scalability Of Sc Culture Through the Use Of Mcsmentioning

confidence: 99%

Microcarriers for Upscaling Cultured Meat Production

2020

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“…Positive for ARS, ALP activity, and osteogenesis-related genes upregulation [62] Not tested Positive for OROS [62] hMSC-hTERT Positive for ARS [64], ALP activity [33], upregulation of osteogenesis-related genes [33,63,68], and in vivo bone formation [63,68] Positive for ABS [64] and ColII immunostaining [63,67] in 2D culture Positive for OROS and upregulation of adipogenesis-related genes [33] TERT4 (hMSC-hTERT derived)…”

Section: Kpmentioning

confidence: 99%

Usefulness of Mesenchymal Cell Lines for Bone and Cartilage Regeneration Research

Piñeiro-Ramil

Sanjurjo‐Rodríguez

Castro-Viñuelas

et al. 2019

IJMS

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The unavailability of sufficient numbers of human primary cells is a major roadblock for in vitro repair of bone and/or cartilage, and for performing disease modelling experiments. Immortalized mesenchymal stromal cells (iMSCs) may be employed as a research tool for avoiding these problems. The purpose of this review was to revise the available literature on the characteristics of the iMSC lines, paying special attention to the maintenance of the phenotype of the primary cells from which they were derived, and whether they are effectively useful for in vitro disease modeling and cell therapy purposes. This review was performed by searching on Web of Science, Scopus, and PubMed databases from 1 January 2015 to 30 September 2019. The keywords used were ALL = (mesenchymal AND ("cell line" OR immortal*) AND (cartilage OR chondrogenesis OR bone OR osteogenesis) AND human). Only original research studies in which a human iMSC line was employed for osteogenesis or chondrogenesis experiments were included. After describing the success of the immortalization protocol, we focused on the iMSCs maintenance of the parental phenotype and multipotency. According to the literature revised, it seems that the maintenance of these characteristics is not guaranteed by immortalization, and that careful selection and validation of clones with particular characteristics is necessary for taking advantage of the full potential of iMSC to be employed in bone and cartilage-related research.

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Microcarrier choice and bead-to-bead transfer for human mesenchymal stem cells in serum-containing and chemically defined media

Cited by 57 publications

References 37 publications

Human Platelet Lysate Improves Bone Forming Potential of Human Progenitor Cells Expanded in Microcarrier-Based Dynamic Culture

Human Platelet Lysate Improves Bone Forming Potential of Human Progenitor Cells Expanded in Microcarrier-Based Dynamic Culture

Microcarriers for Upscaling Cultured Meat Production

Usefulness of Mesenchymal Cell Lines for Bone and Cartilage Regeneration Research

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