Human cell lines for biopharmaceutical manufacturing: history,status, and future perspectives

Biotherapeutic proteins represent a mainstay of treatment for a multitude of conditions, for example, autoimmune disorders, hematologic disorders, hormonal dysregulation, cancers, infectious diseases and genetic disorders. The technologies behind their production have changed substantially since biotherapeutic proteins were first approved in the 1980s. Although most biotherapeutic proteins developed to date have been produced using the mammalian Chinese hamster ovary and murine myeloma (NS0, Sp2/0) cell lines, there has been a recent shift toward the use of human cell lines. One of the most important advantages of using human cell lines for protein production is the greater likelihood that the resulting recombinant protein will bear post-translational modifications (PTMs) that are consistent with those seen on endogenous human proteins. Although other mammalian cell lines can produce PTMs similar to human cells, they also produce non-human PTMs, such as galactose-α1,3-galactose and N-glycolylneuraminic acid, which are potentially immunogenic. In addition, human cell lines are grown easily in a serum-free suspension culture, reproduce rapidly and have efficient protein production. A possible disadvantage of using human cell lines is the potential for human-specific viral contamination, although this risk can be mitigated with multiple viral inactivation or clearance steps. In addition, while human cell lines are currently widely used for biopharmaceutical research, vaccine production and production of some licensed protein therapeutics, there is a relative paucity of clinical experience with human cell lines because they have only recently begun to be used for the manufacture of proteins (compared with other types of cell lines). With additional research investment, human cell lines may be further optimized for routine commercial production of a broader range of biotherapeutic proteins.     

Haramonas dimorpha gen. et sp. nov. (Raphidophyceae), a new marine raphidophyte from Australian mangrove*

A new raphidophyte flagellate is described from the river mouth of the Daintree River, north-east Australia where there are extensive mangrove forests. The organism has two distinct phases: a club-shaped motile form, and a more or less spherical benthic non-motile form. The motile cell is metabolic and possesses 10–20 chloroplasts. The chloroplasts are imbricated, somewhat reminiscent of roofing tiles. A unique structure has been found at the posterior end of the cell. It is a tubular invagination, which consists of a single membrane supported by many small flattened vesicles. In addition, the structure is surrounded by many fibril-containing vesicles. The tubular invagination is different from both the contractile vacuoles and the pusules of dinoflagellates in both behavior and structure. Based on the possession of these unique features, it is concluded that the this mangrove flagellate should be described as a new species in a new genus and the name Haramonas dimorpha Horiguchi gen. et sp. nov. is proposed.     

Heterochromatin and cytogenetic polymorphisms in Cebus apella (Cebidae,Platyrrhini)

Cytogenetic studies have been carried out in 39 specimens of C. apella of different origins. Three different morphologies, one affecting the long arm of chromosome 4 and two affecting pair 17, have been detected. In each case, they can be related by paracentric inversions. Heterochromatin polymorphisms affecting terminal or interstitial C+ regions have also been observed. The length of the terminal heterochromatic region in the long arms of chromosome 11 is variable in C. apella sp., in C. a. paraguayanus and absent in the C. a. nigritus specimens studied. Interstitial C + bands can be observed in the long arms of the biarmed chromosomes 4 and 6, and in the long arms of the acrocentric pairs 12, 13, 17, 18, 19, 20, and 21. Interstitial C + bands in the long arms of chromosomes 4, 12, 17, and 19 are present in all animals studied, although their size is variable, especially in the case of chromosomes 17 and 19. © 1995 Wiley-Liss, Inc.     

Nothacrobeles,n. gen., with Descriptions of Four New Species (Nematoda: Cephalobidae)

A new nematode genus, Nothacrobeles, is proposed in the subfamily Acrobelinae. Four new species are described and one new combination made. The five species exhibit a progression from short-to-long, bifurcate, elaborately fringed labial probolae, the longest probolae resembling those of species of Acrobeles.     

DEUCALION GEN. NOV. AND ANISOSCHIZUS GEN. NOV. (CERAMIACEAE,CERAMIALES), TWO NEW PROPAGULE-FORMING RED ALGAE FROM SOUTHERN AUSTRALIA1

Two new propagule-farming red algae from southern Australia, Deucalion levringii (Lindauer) gen. et comb. nov. and Anisoschizus propaguli gen. et sp. nov., are described and defined largely on their development in laboratory culture. Deucalion is included in the tribe Compsothamnieae on the basis of its subapical procarp and alternate distichous branching. It differs from the other genera included in that tribe in that it produces 3-celled propagules, polysporangia, a subapical cell of the fertile axis which bears 3 pericentral cells, and an apparently post-fertilization involucre which develops from the hypogenous and sub-hypogenous cells of the fertile axis. Its gametophyte morphology has been elucidated in culture, as only sporophytes are known from the field. Gametophytes do not appear to produce propagules. Anisoschizus is provisionally included in the tribe Spermothamnieae on the basis of its subdichotomous branching, possession of a prostrate system and the production of polysporangia. It differs from the other genera of the tribe in the production of 2-celled propagules. Observations on the germination of the “monosporangia” of Mazoyerella arachnoidea and Monosporus spp. indicate that they are analagous to the propagules of Deucalion and Anisoschizus. The nature of these propagules and their role in recycling the parent plant are discussed and contrasted with true monosporangia. It is recommended that Monosporus be maintained as a form genus containing representatives from more than one tribe, as exemplified by plants from Lord Howe I. provisionally identified as M. indicus Boergesen which have both prostrate and erect, as opposed to only erect, axes.     

Determination of fetal biparietal diameter without the use of ultrasound in squirrel monkeys

This study evaluated manual caliper measurement of fetal BPD in Saimiri through the abdomen of the dam (TBPD) for correlation with paired ultrasound measurements, prediction of delivery date, subspecies variation, prediction of pregnancy outcome, and correlation between postpartum BPD and TBPD. Regression analysis revealed a close relationship between TBPD measurements and those obtained by ultrasound (P < 0.001). TBPD for Saimiri boliviensis boliviensis increased from a mean of 14.8 ± 0.3 mm for 11 weeks prepartum to a delivery week measurement of 33.2 ± 0.2mm. Delivery-week TBPD of Saimiri boliviensis peruviensis were significantly smaller than Saimiri sciureus sciureus (P < .05). Delivery week TBPD had a correlation coefficient of 0.64 with paired post-patrum measurements. No complications were associated with the technique. Delivery date could be predicted within two weeks. TBPD is an inexpensive, safe, rapid method of approximating fetal growth during the last half of pregnancy in Saimiri.