Understanding the role of Cripto-1 in cancer progression and therapeutic strategies

Zeng, Qingfang; Gao, Yuzhen; Zhou, Ying

doi:10.1007/s12094-022-03023-2

Cited by 3 publications

(1 citation statement)

References 78 publications

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“…In the present study, we aimed to scale up the production of a therapeutic protein called Cripto-1 using recombinantly modified HEK293 cells grown in PF microcarriers. Cripto-1 is a 27 kDa membrane glycosylphosphatidylinositol (GPI)-anchored protein, which belongs to the EGF–CFC protein family and plays an important regulatory role in embryonic development [ 46 ]. It can act both as a ligand via the Nodal/Alk4-independent signaling pathway and as a co-receptor for Nodal through activation of the ALK4/ActRIIB receptor complex [ 5 , 47 , 48 ].…”

Section: Introductionmentioning

confidence: 99%

Biomanufacturing Recombinantly Expressed Cripto-1 Protein in Anchorage-Dependent Mammalian Cells Growing in Suspension Bioreactors within a Three-Dimensional Hydrogel Microcarrier

Lev

Bar-Am

Lati

et al. 2023

Gels

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Biotherapeutic soluble proteins that are recombinantly expressed in mammalian cells can pose a challenge when biomanufacturing in three-dimensional (3D) suspension culture systems. Herein, we tested a 3D hydrogel microcarrier for a suspension culture of HEK293 cells overexpressing recombinant Cripto-1 protein. Cripto-1 is an extracellular protein that is involved in developmental processes and has recently been reported to have therapeutic effects in alleviating muscle injury and diseases by regulating muscle regeneration through satellite cell progression toward the myogenic lineage. Cripto-overexpressing HEK293 cell lines were cultured in microcarriers made from poly (ethylene glycol)-fibrinogen (PF) hydrogels, which provided the 3D substrate for cell growth and protein production in stirred bioreactors. The PF microcarriers were designed with sufficient strength to resist hydrodynamic deterioration and biodegradation associated with suspension culture in stirred bioreactors for up to 21 days. The yield of purified Cripto-1 obtained using the 3D PF microcarriers was significantly higher than that obtained with a two-dimensional (2D) culture system. The bioactivity of the 3D-produced Cripto-1 was equivalent to commercially available Cripto-1 in terms of an ELISA binding assay, a muscle cell proliferation assay, and a myogenic differentiation assay. Taken together, these data indicate that 3D microcarriers made from PF can be combined with mammalian cell expression systems to improve the biomanufacturing of protein-based therapeutics for muscle injuries.

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